HU200183B - Process for producing beta-lactam antibiotics and pharmaceutical compositions comprising same - Google Patents

Abstract

Antibacterially active and animal growth-promoting novel beta -lactam compounds of the formula <IMAGE> in which R1 represents the radical <IMAGE> Y representing N or CR9, or Y-R7 representing >C=O or >C=N-R7, Z representing O, S, or NR10, and R2 represents hydrogen or a protective group.

Description

The present invention relates to a process for the preparation of β-lactam antibiotics and pharmaceutical compositions containing them. The compounds of the present invention are antibacterial, in particular orally active.

Certain 7a-aminoalkyl cephalosporin derivatives having different substituents at the 3-position of the molecule are known to have antibiotic activity. Examples are cephalexin / 7- (D-o-phenylglycylamido) 3 methyl cephen-4-carboxylic acid, cephalochloro (7- (D - N -phenylglycylamido) -3-chloro-3-cephem-4-carboxylic acid). carboxylic acid / (United Kingdom Patent Nos. 1,174,335 and German Patent Nos. 24,068,698 and 27,28,578).

Antibiotic α-aminopencillanic acids such as azampicillin (British Patent No. 938,321) and amoxicillin (British Patent No. 1,339,605) are also known.

The compounds of the invention are represented by formula (I). In this formula

X is a group of formula (1), (2) or (3) wherein

Hal means halogen,

R is amino benzothiazolyl, benzimidazol-1, 2-amino-6-methyl benzothiazolyl, aminobenzimidazolyl; trifluoromethylbenzimidazolyl, methylbenzimidazolyl, 2,3-dihydro-2-oxo-1H-benzimidazolyl, aminobenzoxazolyl or benzthiazolyl

Due to the presence of an asymmetric carbon atom represented by * in formula 0), the new β-lactam antibiotics include D, L and D.L forms. The D-shape is preferred.

Both diastereomeric mixtures and D- and L-forms of the compounds of the invention are useful for controlling bacterial infections.

Compounds of formula (1) formula is prepared by reacting (II) with a compound of the formula: - wherein ^R1 is as defined above and R ² is an amino-protecting group - the carboxy group such as pivaloyl chloride, chloroformic acid ethyl or isobutyl ester to give the mixed anhydride, or methanesulfonic acid chloride to the mesylate, or, for example, 1-hydroxybenzotriazole or dicyclohexylcarbodiimide to activate the ester of formula 011, wherein X is and then deprotecting.

The coupling of amino acids of formula II, wherein R ¹ and R ² are as defined above, with the β-lactam compounds of formula III, wherein X is as defined above, can be accomplished by cephalosporin and penicillin chemistry is known by a number of processes.

It is preferable to activate the amino acids of formula (O) - wherein R and R ² are as defined above and then couple with the β-lactam compounds of general formula (IH) dissolved in the form of the amino salts, wherein X is as defined above, Activation is particularly preferably carried out with pivalic acid chloride 25 or the sulfonic acid derivative of formula 0V to give compounds of formulas Va and Vb. The reaction scheme is illustrated in Schemes a and b. In the formulas

R ^ is as defined,

R ² is an amino protecting group,

T is R ¹ -SO 2 -O or -alogen and

R ¹³ is 1-10 optionally substituted with fluoro or chloro, cyano, phenyl, C 1-4 alkyl, C 1-4 alkyl, C 1-4 alkoxy, or C 1-4 alkylthio C 1 -C 4 alkyl, or optionally with fluorine, chlorine or bromine, cyano, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, C 1 -C 4 alkylthio, C 1 -C 4 alkylcarbonyl, nitro or phenyl substituted with trifluoromethyl.

When R ³ is substituted, it preferably contains from 1 to 3 of the listed substituents.

Preferably R is methyl or p-tolyl.

The mixed anhydrides of formula (Va) may be prepared by reacting an acid of formula (III)

1.4 equivalents of the amine are dissolved in a solvent and then reacted with 1-1.2 equivalents of the sulfonic acid derivative of the formula 0V).

As the solvent, any solvent which is stable under the reaction conditions can be used. Examples include diethyl ether, tetrahydrofuran, acetonitrile, acetone, methylene chloride, chloroform and dimethylformamide.

Suitable amines are tertiary amines such as triethylamine and tributylamine, as well as spherically inhibited amines such as diisopropylamine.

The reaction may be carried out at a temperature between -80 'C and room temperature, preferably between -60' and 0 '. The activation of Q-SO2-CH3 in dimethylformamide is preferably carried out at a temperature between -40 'C and -60' C for 0.2-24 hours, preferably 0.5-5 hours.

The solvents listed in the preparation of compounds of formula (V), for example, the amines mentioned therein, may be used to dissolve the compounds of formula (IH) wherein X is as defined above.

Activation of the amines of formula (Π) is particularly preferably carried out by preparing the activated esters, for example, with N-hydroxysuccinimide and dicyclohexylcarbodiimide or 1-hydroxybenzotriazole and dicyclohexylcarbodiimide.

Suitable solvents are any of the solvents listed above for the preparation of compounds of formula (V).

The reactions may be carried out at temperatures between -30 'C and * 100' C. Preferably, the activation is carried out with hydroxybenzotriazole and dicyclohexyl cartleclimide in dimethylformamide at room temperature for 2 to 6 hours, then the excellent dicyclohexylurea is suctioned off and reacted for 2-24 hours with a solution of the amine salt of compound 011. To dissolve compounds of formula (0), compounds of formula (V)

The solvents listed in the preparation of -2EN 200183Β and the amines listed therein may be used as the base.

Amino and carboxyl protecting groups and carboxyl activation are described in M. Bodanszky, Principles of Peptide Synthesis, Springer-Verlag, 1984. E. Gross, J. Meienhofer, The Peptides Vol. 2, Academic Press, 1980. .

Stereochemically uniform D- and L-forms of the compounds of formula I are prepared by separating the diastereomeric mixture, for example, on a Merck, DuPont or Whatmann high performance liquid chromatography column.

The pure D- or L-form (preferably the D-form) may also be prepared by chemical separation of racemate with, for example, dehydroabiethylamine, phenylethylamine or camphorsulfonic acid in step 2 of the racemic amino acid formula Racemate separation is carried out on an N-acetylamino acid derivative such as subtilisin, penicillinacylase or porcine kidney acylase, and the resulting stereochemically uniform D- or L-shaped compound of formula (Π) is further converted as indicated.

Compounds of formula II, wherein R ¹ and R ² are as defined above, are only partially known. Compounds of formula (II), wherein R ¹ and R ² are as defined above, are known in the art, such as The most important key compounds in the preparation of compounds of formula (U) in the reaction are compounds of formula (VI): wherein R ¹ is as defined above and R ¹⁴ is C 1 -C 4 alkyl.

Reduction of the esters to diisobutylaluminum hydride (DIBAL) and sodium bis (2-methoxyethoxy) aluminum hydride (Red-Al) (step 1) is known in the art from E. Winterfeld, Synthesis 1975.617; A. E. G. Miller et al.

J. Org. Chem. 24, 627 (1959).

The oxidation of primary alcohols with manganese (IV) oxide or pyridinium chlorochromate to the aldehyde (step 2) is also known in the art: Methoden Der Organischen Chemie, Houben-Weyl, Bd. 4/1 b; G. Piancatelli et al., Synthesis 1982, 2445.

The new amino acids of formula Hb, wherein R ¹ is as defined above, are prepared by reacting the aldehyde with sodium cyanide and ammonium carbonate according to a method known in the art (E. Ware, Chemical Reviews 46, 403 (1950)). (step 3) followed by 10% sodium hydroxide solution, 48% hydrobromic acid solution, barium hydroxide solution or lithium hydroxide solution (step 4).

The following describes, by way of example, the preparation of some new amino acids and their precursors. Schemes 1 to 6 wherein R ¹⁴ is as defined above and is hydrogen, straight or branched or cyclic, saturated or unsaturated, optionally with halogen, hydroxy, C 1-6 alkoxy, C 1-6 alkoxycarbonyl, cyano, carboxyl, optionally substituted aryl, SOjH or optionally substituted amino substituted with up to 10 carbon atoms or optionally substituted aryl and can be halogen, C 1-8 alkoxy or C 1-8 alkylthio, optionally substituted amino, SO 2 - (C 1 -C 8) alkyl or PO (OH) 2, SH, OH, S-phenyl or O-phenyl, guanidino, amidino; - NHNH2 or NHOH group, an optionally substituted heterocyclic or S-heterocyclic group or Oheterocikhisos Csöpi ° ^rt l0

R is hydrogen, hydroxy or amino, or straight or branched chain cyclic, saturated or unsaturated, optionally substituted with up to 10 carbon atoms, optionally substituted with halogen, optionally substituted amino, hydroxy, cyano or C 6-10 aryl C 6 -C 10 aryl, or

R ⁹ and R ¹⁰ together form an optionally interrupted C 2 -C 4 methylene chain with oxygen or sulfur;

R ¹¹ and R ² are the same or different and are selected from the group consisting of hydrogen, aryl, preferably phenyl, C 1-8 alkyl, preferably C 1-5 alkyl, C 7-14 aralkyl, preferably benzyl or C 2-10, preferably acetyl or benzoyl.

1) Preparation of benzothiazolylglycine derivatives

The starting compounds of the substituted benzothiazole glycine moieties are the 0-, m- or p-aminobenzoic acid esters. Benzothiazole carboxylic acid derivatives are prepared, for example, according to Scheme 2 (J.46 Wood, Substitution and Addition Reactions of Thiocyanogen, Organic Reactions, Vol. 240 (1946); Bogert T. et al., J. Am. Soc., 47, 3078 (1925); Bogert T., J. Am. Chem. Soc., 52, 1529 (1935); J.M. Spraque et al., Thiazoles and Benzothiazoles, Heterocyclic Compounds, Vol. J. Wiley & Sons). The starting compounds of the substituted benzothiazolylglycine derivatives may also be the α-amino-α- (p-aminophenyl) acetic acid esters. This reaction is illustrated in Scheme 3.

2) Preparation of benzimidazole / glycine derivatives

Examples of substituted benzimidazole-1-glycine derivatives include 3,4-diaminobenzoic acid derivatives. Substituted benzimidazole carboxylic acid esters are prepared according to Scheme 4. (Pr. N. Preston, 1974, 74, 279; R. Rastogi et al., Synthesis 1983, 861; PN Preston, Benzimidazoles and Congeneric Tricyclic Compounds, Vol. 40, I and,, J. Wiley & Sons (1980, 1981)).

3. Preparation of benzoxazolylglycine derivatives

The starting compounds for the preparation of the substituted benzoxazolylglycine derivatives include 3-amino-4-hydroxybenzoic acid, 3-hydroxy-4-aminobenzoic acid and Da-amino- (3-amino-4-hydroxy-phenyl) - acetic acid methyl ester. The novel anellated phenylglycine derivatives and substituted benzoxazole carboxylic acid derivatives are prepared, for example, according to Scheme 5 (J.W. Cornforth, Benzoxazoles and Related Systems, Heterocyclic Compounds, 5).

413, J. Wiley & Sons (1957); R. Lakham et al. Advances in Oxazole Chemistry, Advances in Heterocyclic Chemistry, Vol. 17, 99, Academic Press (1974).

4) Preparation of benzthiazolyl and benzthiadiazolylglycine derivatives.

The starting materials for the synthesis of substituted benzthiazolyl and benzthiadiazolylglycine derivatives are 3,4-diaminobenzoic acid and ethyl p-aminobenzoic acid. The novel benzene ring-fused carboxylic acids or carboxylic acid esters are prepared, for example, according to Scheme 6 (J.B. Carr, J. Heterocyclic Chem. 2. 1149 (1972)).

The penicillin and cephalosporin-based compounds described below are used to prepare compounds of formula I wherein R ³ , R ⁴ and R ⁵ are as defined above.

1) Cephalosporin compounds of formula (IHa)

These include 7-amino-3-methyl-3-cephem-4-carboxylic acid (7-ADCA) and 7-amino-3-chloro-3-cephem-4-carboxylic acid (7-ACCA), which are described in J. Med. Chem. 12,310 CW Ryan et al. (1969), U.S. Patent 3,994,884. United States Patent No. 2,606,196; German Patent Application Laid-Open No. Among the compounds of formula (a), the following are listed:

R - CH ₃ , Cl, H, Br, OCH 3, CH - CH 2, CH - CHCH 3 (cis).

2) Penicillin compounds of formula (IHb)

In addition to 6-aminopenicillanic acids, 6-amino-bisnorpenicillanic acids (British Patent No. 1,546,622) and 6-α-formamidopenicillins (PH Milner et al. (J. Chem. Soc. Chem. Commun. 1984, 1335).

The compounds of formula (I) according to the invention, wherein R ¹ and X are as defined above, exhibit a broad spectrum of Gram-positive and Gram-negative bacteria, especially

Against Staphylococci, Streptococcus, Enterococcus and Haemophilus influenzae.

The compounds of the present invention have very good activity against various microorganisms, such as Staphylococci, such as Staph. aureus, Staphl. epidermidis, Streptococcus mites such as Streptococcus pyogenes, Streptococcus faecalls, Enterobak20 terriers such as Escherichia coli, Klebsiella, Salmonella, Shigella, Proteus varieties such as Proteus mirabilis opposite.

Based on their properties above, the compounds are useful as chemotherapeutic agents in human and veterinary medicine. The following table shows the minimum inhibitory concentration (MHC values, mg / ml) of the compounds of the invention and of Cafaclor. / M. Gorman et al., Cefachlor, Chronicles of Drug Discovery, Volume 2, 49,

J. Wiley & Sons (1983) /.

MHK values were determined by agar dilution method after solid cultivation for 18-24 hours at 37 ° C using a Multipoint inoculator. The culture medium used is Isosensitest-agar medium.

The number of the production example

The bacterium 2a 8 12a 15a 20 22 23 cefaclor E. coli T2 32 4 4 128 16 16 128 8 E. coli A 261 32 4 4 128 16 16 128 2 E. coli Ncum. 8 1 2 8 4 2 4 2 Ecoli 183/58 > 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 E. coli F14 128 32 128 > 128 128 > 128 256 16 E. coli C165 24 0.5 2 82 4 3 2 1 E. coli 4322 4 0.5 16 8 2 4 4 1 Klebs 57USA 32 4 4 64 16 16 32 8 Klebs 63 1 0.5 1 8 • 2 1 2 1 Klebs 1852 128 8 8 128 64 64 128 8 Klebs 6097 2 0.5 1 16 2 2 2 2 Serratia 16001 > 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 Serratia 16002 > 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 Provid 12012 > 128 32 32 > 128 > 128 128 > 256 32 Prot. Norg. 932 126 > 128 > 128 > 128 > 128 > 128 32 > 128 Prot. Volg. 9023 128 64 > 128 128 > 128 128 > 256 64 Prot.volg. 1017 > 128 > 128 > 128 > 128 > 128 > 128 128 > 128 Prot. Volg. Woman > 128 > 128 > 128 > 128 > 128 > 128 4 > 128 Prot. meadow 10007> 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 Prot.mir. 1235. 1 2 4 8 8 2 4 4 Staph. 1756 128 128 128 > 128 128 128 > 256 > 128 Staph. 133 03 0.5 2 4 2 1 1 2 Staph. 25022 03 0.5 2 4 4 1 2 4

-4HU 200183Β

8

The number of the production example

Λ bacteria 2a 8 12a 15a 20 22 23 cefaloram Staph. 25 470 128 64 > 128 > 128 128 > 128 > 256 > 128 Staph. E. 25185 0.06 0,125 64 05 8 0.25 2 16 Enterococc 9790 8 32 32 64 32 64 > 256 64 Entenx23 ± 58 2 16 32 32 8 64 > 256 32 Psdm. F41 > 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 Psdm. Walter > 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 Psdm. 7035 > 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 Psdm. 7451 > 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 Enercb. cl. 5605 > 128 > 128 > 128 > 128 > 128 > 128 > 256 > 128 Enercb. cl. 5744 128 32 8 64 64 16 > 256 8 Achroroh 2005 0:25 0.5 <0.06 2 0.25 0.5 0.5 0.25

MHK values for orally active cephaloeporn derivatives

Ideal and / or systemic diseases caused by the above pathogens or their spots can be treated and / or prevented using the compounds of the present invention. 5

The compounds of the present invention can be used to ameliorate and / or cure diseases such as respiratory and pharyngeal diseases such as Otitis, Pharyngitis, Pneumonia, Peritonitis, Pyelonephritis, Cystitis, Endocarditis, Systemic Infections, Bronchitis, Arthritis,

The pharmaceutical compositions of the present invention contain one or more compounds of the present invention in addition to non-toxic inert pharmaceutically acceptable carriers.

The compositions of the invention may also be presented in unit dosage forms. This means that the compositions may be prepared, for example, in the form of tablets, dragees, capsules, pills or ampoules, each containing a fraction or multiple of the active ingredient. Dosage units contain, for example, a unit dose of 1,2,3 or 4 units or a unit dose of 1 / 2,1 / 3 or 1/4 units. Preferably, the unit dose contains an amount of the active ingredient which is administered in a single application and which generally corresponds to the total amount, half, third, or quarter of the total daily dose. 30

Non-toxic inert pharmaceutically acceptable carriers include solid, semi-solid, or liquid diluents. Fillers and formulation aids are known.

Preferred pharmaceutical compositions include tablets, dragees, capsules, pills, granules, suppositories, solutions, suspensions and emulsions, pastes, ointments, gels, creams, solutions, powders, and sprays.

The tablets, dragees, capsules, pills and granules may contain, in addition to the active ingredient (s), the customary carriers. These include (a) fillers such as starch, lactose, raw sugar, glucose, mannitol and silica, (b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, (c) humectants such as glycerol, (d) disintegrants such as agar, calcium carbonate and sodium carbonate, (e) solubilizers such as paraffin, absorption enhancers such as kwlferner ammonium compounds, (g) wetting agents such as cetyl alcohol, glycerol monranearate, (h) adsorption agents such as kaoMh and bentonite, and (i) lubricants, such as talc, calcium and magnesium stearates, solid polyethylene glycols or mixtures of the substances listed in (a) - (ft) Oatt.

The tablets, troches, capsules, pills and granules may be provided with a conventional coating or coating, optionally containing an opacifying agent, and may be prepared so that the active ingredient (s) is / are they may be delayed, for example in the case of polymers or waxes.

The active compounds) may optionally be formulated in microencapsulated form with one or more specified carriers.

The suppositories may contain, in addition to the active ingredient (s), the customary water-soluble or water-insoluble carriers, such as polyethylene glycols, fats such as cocoa butter and long chain esters (e.g.

Ointments, pastes, creams and gels may contain, in addition to the active compounds, the customary carriers. Examples include animal and vegetable oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicones, bentonite, silica, talc and zinc oxide, or mixtures thereof.

Powders and sprays may contain the usual carrier (s) in addition to the active ingredient (s). Examples are lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powders and mixtures thereof. In addition, the sprays may also contain a propellant such as chlorofluorocarbons.

The solutions and emulsions may contain, in addition to the active compounds, the customary carriers.

-5HU 200183Β

These include solvents, antisolvents and emulsifiers such as water, ethyl alcohol, isopropyl alcohol, benzyl benzoate. propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, especially cottonseed oil, castor oil and sesame oil, glycerol, glycerol forms, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters or mixtures thereof.

For parenteral administration, solutions and emulsions may also be formulated in sterile or blood isotonic form.

The suspensions may contain, in addition to the active ingredient (s), customary carriers such as liquid diluents such as water, ethyl alcohol, propylene glycol, suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystals. cellulose, aluminum metahydroxide, bentonite, agar and tragacanth or mixtures thereof.

The formulations listed above may contain coloring agents, preservatives, and flavoring agents such as peppermint oil or eucalyptus oil, and a sweetening agent such as saccharin.

The compositions of the present invention preferably contain from 0.1 to 10

995, preferably 0.5-95% by weight,

In addition to the compounds of the invention, the compositions of the present invention may contain additional pharmaceutical active ingredients.

The pharmaceutical compositions of the invention may be administered topically, orally, parenterally, intraperitoneally and / or rectally, preferably orally or parenterally, such as intravenously or intramuscularly.

In both veterinary and human medicine, it has been found advantageous to administer the compounds of the invention in a total amount of 5 to 1000, preferably 10 to 200 mg / kg body weight, every 24 to 40 hours, optionally in multiple unit doses.

The unit dose of the active ingredient (s) is preferably 1-

It is present in an amount of 250, preferably 3-60 mg / kg body weight. In some cases, it may be necessary to deviate from these dosage amounts, depending on the patient, his weight, the nature and severity of the disease, the mode of administration of the composition and the drug, and the duration and interval of administration. Thus, in some cases, it may be sufficient to use less than the stated amount of active ingredient, while in other cases, using more than this amount. Determining the optimum dosage and dosage regimen required at any time is within the skill of the art. 55

When the compounds of the present invention are used as feed rations, the active ingredient is administered in the usual concentrations and formulations together with the feed or the feed composition or the drinking water. Thus, 60 infections caused by Gram-negative or Gram-positive bacteria can be prevented, cured and / or cured while at the same time improving growth and marketing of feed. 65

The novel compounds of the present invention have potent antibacterial activity which has been tested both in vivo and in vitro. In addition, the compounds are well absorbed orally

The active compounds of the present invention may be combined with other antimicrobial agents and lactamase inhibitors, such as penicillinase-resistant penicillins and clavulanic acid or aminoglycoside antibiotics such as gentamycin, isomycin, or canamycin, to broaden or broaden the spectrum of action.

The invention is illustrated by the following embodiments.

Example 1

Preparation of DL-7- (2-Aminobenzothiazol-2-yl-glycylamido) 3-chloro-3-cephem-4-carboxylic acid (I-1)

a) Preparation of 2-Amino-6- (hydroxymethyl) -benzothiazole (la) Compound 6-ethyl ester of 2-aminobenzothiazole carboxylic acid (0.225 mol) was suspended in 1000 ml of tetrahydrofuran and cooled to -70 ° C. 5625 ml (0.675 mol) of diisobutylaluminum hydride (DIBAL, 20% toluene solution, 1.2 mol) were added slowly dropwise to the reaction temperature and under a nitrogen atmosphere. The reaction mixture was allowed to stand overnight at -70 ° C to -40 ° C and stirred for 3 hours / without cooling. The resulting reaction mixture was cooled to -70 ° C, water (61.5 mL) was added dropwise (strongly exothermic) and the resulting mixture was allowed to warm to room temperature. To the resulting reaction mixture was added 305 mL of saturated brine and stirred for 1 hour at 20 ° C. The precipitated aluminum hydroxide is filtered off with suction, washed with tetrahydrofuran and the filtrate is evaporated to dryness (36 g). The residue was dissolved in hot ethanol, the insolubles were filtered off and the filtrate was evaporated to dryness. Yield: 255 g (63%).

CeH ₈ N ₂ OS (180.2)

NMR (DMSO): δ 4.5 (d, 2H), 5.13 (t, 1H), 7.18 (d, 1H), 7.3 (d, 1H), 7.4 (s, 2H) ), 7.59 (s, 1H) ppm.

b) Preparation of 2-aminobenzothiazole-6-carboxaldehyde (1b)

Compound (Ia) (110.5 g, 0.613 mol) in THF (200 mL) was stirred with manganese (IV) oxide (319.8 g, 3.678 mol) for 3 days at room temperature. The reaction mixture was filtered and the residue was washed with tetrahydrofuran and the filtrate was concentrated in vacuo. Yield: 74 g (68%).

C8H6N ₂ OS (178.1)

NMR (DMSO) δ- 7.48 (d, J-7.5Hz, 1H), 7.8 (d, J-7.5Hz, 1H), 8.08 (s, 2H), 8.26 (s, 1H), 9.92 (s, 1H) ppm.

c) Preparation of 5- (2-Aminobenzothiazol-6-yl) -2,4-imidazolidinedione (Compound (Ic))

A solution of (1b) (22.5 g, 0.126 mol) in methanol (400 mL) was added dropwise to a solution of ammonium carbonate (50.9 g, 0.536 mol) and sodium cyanide (9.6 g, 0.196 mol) in water (400 mL). the resulting reaction mixture was stirred for 20 hours

Stir at -63 ° C. The methanol was evaporated in vacuo and the residue was acidified to pH = 2 with 2N hydrochloric acid at 0 ° C, then the pH was adjusted to 4 with 2N sodium hydroxide solution and the precipitated product was filtered off with suction. Yield: 20.5 g (65%).

Elemental analysis for C10H8N4O2S (248.3):

Found: C, 48.38; H, 3.25; N, 22.56; S, 12.91. Found: C, 48.4; H, 3.5; N, 21.2; 3%.

NMR (DMSO): δ: 5.17 (s, 1H), 7.17 (q, 1H), 7.35 (d, 1H), 7.57 (s, 1H), 7.62 (s, 1H) ), 8.4 (s, 1H), 10.610.82 (bs, 1H) ppm.

d) Preparation of DL-α-Amino-α- (2-aminobenzothiazol-6-yl) acetic acid (DL-6-aminobenzothiazolylglycine, Compound (1d)) Preparation of Compound (Le) in g (0.081 mol). Lithium hydroxide (19.4 g, 0.81 mol) in water (400 ml) was heated with stirring at 100 ° C for 24 hours. The resulting solution is filtered hot, the residue is washed with hot water and the filtrate is acidified to pH 2 with concentrated hydrochloric acid while cooling with ice. The resulting reaction mixture was stirred for 15 minutes at 0 ° C and the resulting solution containing the precipitated material was brought back to pH 4.5. The resulting precipitate was filtered off with suction, washed with water / acetone for a year and dried in vacuo. Yield: 10.4 g (56%).

Analysis for C9H9N3O3S x 2.1 _H2 O (232.3) Calc'd:

Calculated: C, 45.05; H, 4.20; N, 17.51; S, 23.36. Found: C, 45.3; H, 4.50; N, 15.50; S, 13 40%.

NMR (DMSO): δ: 5.11 (s, 1H), 7.8 (s, 2H), 8.11 ($, 1H) ppm.

e) DL-α- (tert-Butyloxy) carbonylamino-α- (2'-tert-butyloxyZ-carbonylamino-benzothiazol-1-yl) acetic acid (di-Boc derivative) and DL- Preparation of α- (tert-butyloxy) carbonylamino-α- (2-aminobenzothiazol-6-yl) acetic acid (mono-Boc derivative) / (le)

Compound (1d) (20.9 g, 0.0896 mol) was suspended in a mixture of water (100 ml) and dioxane (130 ml) and dissolved in 140 ml of 2N sodium hydroxide solution. To the resulting reaction mixture was added dropwise di-tert-butyl dicarbonate (78.1 g, 0.358 mole) over 30 minutes and the resulting mixture was stirred overnight. The dioxane was distilled off and the resulting solution was diluted with water and washed with ethyl acetate / petroleum ether. acidified to pH 2 with 2N hydrochloric acid and extracted twice with ethyl acetate / tetrahydrofuran (1: 1), washed with brine, dried over sodium sulfate and evaporated to give the mono- and di-Boc. Yield: 27.4 g.

The product obtained is chromatographed on a silica gel column (Merck 0.04-0.063 mm) using the following solvents and solvent mixtures: 1. 2000 ml methylene chloride, 2. 4000 ml methylene chloride / methanol (10: 0.5); 3. methylene chloride / methanol (10: 1), 4. methylene chloride / methanol (1: 1).

Di-Boc derivative (compound 1e):

Yield: 8.7 g (26%).

C19H25N3O6S (423.5)

NMR (DMSO): δ: 1.37 (s, 9H), 1.52 (s, 9H), 5.12 (d, 1H), 7.45 (d, 2H), 7.65 (d, J -7.5Hz, 1H), 7.96 (s, 1H) ppm.

Mono-Boc derivative

Yield: 9.2 g (32%).

C14H17N3O4S (323.4)

NMR (DMSO): δ 1.36 (s, 9H), 4.98 (d, 1H),

7.3 (q, 2H), 7.48 (s, 2H), 7.54 (s, 1H) ppm.

f) DL-7- [2- (tert-Butyloxy-carbonylamino) -2- (2-tert-butyloxy-carbonylamino-benzothiazol-6-yl) -acetamido] -3-chloro Preparation of -3-cephem-4-carboxylic acid (1f)

Activation of precursor acid:

Compound (le) (5.1 g, 12 mmol) was dissolved in dimethylformamide (40 mL), treated with triethylamine (1.69 mL, 12 mmol), and pivaloyl chloride (1.48 mL, 12 mmol) was added dropwise at -40 ° C. the resulting mixture was stirred for 3 hours at -30 to -15 ° C

Preparation of the amine component:

7-Amino-3-chloro-3-cephem-4-carboxylic acid (7-ACCA) (2.83 g, 12 mmol) was suspended in a mixture of tetrahydrofuran (20 mL) and water (10 mL) and diluted to pH 8.3 with concentrated triethylamine. To the reaction mixture was added 5 ml of dimethylformamide to give a single-phase reaction mixture.

Switching and isolation:

The resulting 7-ACCA solution was sprayed at -40 ° C with the resulting anhydride and stirred in a cooling bath. After 1 hour, 10-20 mL of water was added to the reaction mixture and the pH was adjusted to 7.3 with 10% triethylamine / tetrahydrofuran. .

After an additional two hours, water (200 mL) was added to the reaction mixture, stirred with ethyl acetate / tetrahydrofuran (2: 1) and acidified to pH 1.7 at 0 ° C. The organic layer was separated, washed with brine, dried, The resulting product was filtered off with suction and dried to yield 6.2 g (81%) of the desired product (66%) by HPLC.

C26H30ClN5O8S2 (640.1)

1 H NMR (90 MHz, CDCl 3, TMS, ppm): 1.39 (s, 9H). 1.52 (s, 9H), 3.5-4.02 (br m, 2H), 5.1-5.24 (dd, 1H), 5.38 (d, 1H), 5.62-5.8 (dd, 1H), 7.42-7.53 (m, 1H), 7.68 (d, 1H), 7.98 (s, 1H) ppm.

g) Preparation of DL-7- (2-Aminobenzothiazol-6-yl-glycyl-amido) -3-chloro-3-cephem-4-carboxylic acid (Compound (Ig))

Compound (1f) (8.4 g, 13.1 mmol) was dissolved in methylene chloride (50 mL), treated with anisole (0.5 mL) and trifluoroacetic acid (TFE) (50 mL) at 0 ° C and stirred for 15 minutes without cooling. Acetic acid / methylene chloride was distilled off in vacuo and the resulting oily residue was mixed with ether. The trifluoroacetate was filtered off with suction, washed with ether and dried. Yield: 6.5 g (89%).

C16H14CIN5O4S2 x CF3COOH (553.9)

The product was subjected to high performance liquid chromatography (Hibar 250-4.RP8,10pjn.254nm, 3ml / min. Running time: 1000 ml water / 40 ml acetonitrile / 1 TFE)

Determined content of Retention: 1.10 (ld, 10.8%), 3.08 (L-form, 46.5%), 3.82 (D-form, 38.5%).

24lélda

Preparation of D-7- (2-Aminobenzothiazol-6-yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid and L-form of the compound

A) Separation by chromatography

Compound 2.3 g (Ig) was dissolved in a mixture of 50 ml of runner (1000 ml of water / 50 ml of acetonitrile / 1 ml of TFE) and 4 drops of TFE, stirred for 20 minutes, the insoluble material was suctioned off and the filtrate was filtered again (Millex-GS , 0.22 pm, millipore) and 10 ml portions (460 mg) are slowly pumped into a preparative column (Hibar 250-25, RP 18.7 pm, flow rate 15 ml / min) for 10-15 minutes.

D-shape (vertex Π):

Yield: 350 mg of trifluoroacetate A total of 4.3 g of trifluoroacetate was dissolved in 500 ml of water and applied to a 25 ml column of Amberlite IRA-68 (acetate form) and eluted with 1.5 liters of water (distilled). The eluate was evaporated to a volume of 1000 ml and suction filtered and lyophilized to 0.45 µm (> 1 hour).

Yield: 3.0 g (75%).

Elemental Analysis for C16H14ClO5O4S2 x 3 H2O x 0.5 CH3COOH (523.9). H, 4.23; N, 13.36 Found: C, 38.97;

S, 12.24; Cl, 6.76. Found: C, 38.9; H, 4.0; N, 13.6;

S, 12.0.0: 6.8%.

The product was determined to be 97.4% by HPLC (Hibar 250-4, RP 8.10 µm, 254 nm, 3 mL / prc, 0.5 mg / mL, Rt: 1000 mL water / 30 mL CH 3 COOH).

NMR (DCOOD): δ: 3.58 (d, J-18Hz, 1H), 3.94 (d, J-18Hz, 1H), 5.32 (d, J-5Hz, 1H), 5.75 ( s, 1H), 5.94 (d, J-5Hz, 1H), 7.84 (s, 2H), 8.12 (s, 1H) ppm.

L-shape (vertex I):

Yield: 400 mg trifluoroacetate.

A total of 2.01 g of trifluoroacetate is dissolved in 500 ml of water, applied to a 30 ml column of Amberlite IRA-68 (acetate form) and eluted with 1.5 l of water (distilled). The eluate was concentrated to a volume of 1000 ml and lyophilized. Yield: 1.08 g (60%).

Elemental analysis for C18H14ClN5O4S2 x 3H2O (493.9):

Found: C, 38.90; H, 4.08; N, 14.17. Found: C, 38.6; H, 3.7; N, 13.3.

Content of product by high pressure liquid chromatography (Hibar 250-4, RP 8, 10 µm, 254 nm, 1.8 ml / min, running medium: 930 ml H2O / 20 ml pH7 buffer / 50 ml acetonitrile): 943%

NMR (DCOOD): δ-3.78 (d, J-17.5Hz, 1H), 4.03 (d, J-17.5Hz, 1H), 5.4 (d, J-5Hz, 1H), δ, Δ (s, 1H), 5.92 (d, J-5Hz, 1H). 7.87 (s, 2H), 8.2 (s, 1H) ppm.

B) Preparation of D-phenylglycine as a starting material for the preparation of D-7- (2-aminobenzothiazol-6-yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid

a) Da-Acctamido-a-phenyl-acetic acid / (2a) Compound / Preparation of a 500 g (331 mol) of D-Amino-a-phenyl-acetic acid was suspended in 6 liters of water, cooled to 10 ° C hőmcr ^14, stopping, and 1000 ml NaOH (132 g, 3.31 mol). After 15 min, 675 g (6.62 moles) of acetic anhydride was added to the resulting clear solution at 0 ° C, followed by 397 g (9.93 moles) of sodium hydroxide solution in 1000 ml of water. At this time, the reaction temperature rises from 0 ° C to 30 ° C. The solution is stirred for 20 minutes at pH 9-10 in an ice / brine bath and adjusted to pH 1 with concentrated hydrochloric acid (about 1 liter). The resulting mixture was stirred for 10 minutes, the slurry cooled to 10 ° C was filtered off with suction and washed with water (10 L). The product obtained is dried over potassium hydroxide in vacuo. Yield: 553 g (87%).

C10H11NO3 (193.2)

M.p. 190-191 'C (a) 589-218,6' (c-1,

C2H5OH)

b) Preparation of isomeric mixture of D-α-Acetamido-α- (4-nitrophenyl) acetic acid and D-α-acetamido-α- (3-nitrophenyl) acetic acid (2b)

(2a) (227 g, 1.17 mol) was added slowly at 40-50 ° C to 570 mL of concentrated sulfuric acid. After the addition was complete, the reaction mixture was stirred for 5 minutes at 40 'C and then cooled to 0' C. At -5 ° C to 0 ° C, 115 ml (2.39 mol) of 95% nitric acid solution (d-1.5) is slowly added dropwise over 30-45 minutes. The resulting reaction mixture is stirred for 30 minutes at -5 'C to -10' C, poured onto 8 liters of ice, filtered off with suction and rinsed with plenty of water. Yield: 276.1 g (98%).

C10H10N2O3 (139.2)

M.p. 193.9 'C [a] 589- -193.9' (c-1, C 2 H 5 OH)

c) Preparation of D-α-Acetamido-α- (4-aminophenyl) acetic acid (2c)

240 g (1.0 mole) of the mixture of isomers (2b) were dissolved in 3 liters of 95% ethanol and hydrogenated in the presence of 10 g of palladium on charcoal (10%) for 2 hours at 23 ° C and 10 at. It is mixed with 2.4 liters of water, refluxed and suction filtered through silica gel. After standing overnight, his hair precipitates a fine crystalline yellow material which is filtered off with suction, washed with a little ethanol / water (3: 1) and dried. Yield: 67.2 g

C 10 H 12 N 2 O 3 (208.2).

Mp: 208-211 'C.

M.p .: 190-191 '[i] ^{2589 to 186,} · <C = 1, lnsósavoldat).

NMR (DMSO): δ-1.8 (s, 3H), 5.05 (d, 1H), 632 (d, J-8.5Hz, 2H), 7.02 (d, J-8.5Hz), 2H), 8.35 (d, 1H) ppm.

d) Preparation of D-α-Acetamido-α- (4-aminophenyl) -acetic acid methyl ester (2d)

(2c) (67.7 g, 0.33 mol) was suspended in methanol (400 mL) and lionyl chloride (34.9 mL, 0.487 mol) was added dropwise at -10 ° C to -5 ° C with stirring. a solution is formed. The reaction mixture was allowed to warm to 25 ° C overnight and then evaporated to dryness in vacuo. The residue was partitioned between 1000 mL of ethyl acetate and 800 mL of 10% sodium bicarbonate solution and adjusted to pH 7.5 with 2N sodium hydroxide ol-8HG 200183. The pre-acetate phase is separated off, the aqueous phase is washed twice more with ethyl acetate and the combined extracts are washed with brine. After drying and evaporation of the solvent, 48.1 g (67%) of product are obtained.

C11H14N2O3 (222.2) [a] ² 589- -222.2 ° (c = 1, C2H5OH)

NMR (DMSO): δ-1.85 (s, 3H), 3.55 (s, 3H), 5.1 (d, 1H), 5.15 (s, 2H), 6.52 (d, J -9Hz, 2H), 6.97 (d, J-9H, 2H), 8.45 (d, 1H) ppm.

e) Preparation of D-α-Acetamido-α- (2-aminobenzothiazol-6-yl) acetic acid methyl ester (2e)

(2d) (47.8 g, 0.206 mol) was suspended in glacial acetic acid (620 ml) and solid potassium rhodanide (80 g, 0.823 mol) was added. The reaction mixture was stirred at room temperature for 45 minutes to give a clear solution which was cooled to 10 ° C and treated dropwise with 11.6 mL (0.226 mol) of glacial acetic acid in 100 mL of glacial acetic acid over 15 minutes. The resulting reaction mixture was stirred at 10-15 ° C for one hour.

The resulting crystalline material is distilled off in glacial acetic acid under vacuum, the residue is suspended in 1000 ml of water and adjusted to pH 6.3 with sodium carbonate. The resulting reaction mixture was stirred at 70-80 ° C for 2 hours, extracted several times with ethyl acetate, and the combined ethyl acetate layer was washed with brine and dried over sodium sulfate. Yield:

33.8 g (59%).

Elemental analysis for C12H13N3O3S (279.3):

H, 4.7; N, 15.0; S, 11.5. Found: C, 52.0; H, 5.5; N, 14.2; S, 11.0. %.

M.p. 190-191 'C fa] 589- -126,0' (c-1, C 2 H 5 OH)

NMR (DMSO): δ-2.19 (s, 3H), 3.62 (s, 3H). 5.35 (d, 1H), 7.2 (d, 1H), 7.33 (d, 1H), 7.54 (s, 2H), 7.65 (d, 1H), 8.67 (d) , 1H) ppm.

f) Preparation of Da-Amino-α- (2-aminobenzothiazol-6-yl) acetic acid dihydrochloride (2f) g (0.251 mol) (2e) for 40 minutes at 40 ° C in 700 ml of 6N The resulting clear solution was stirred for an additional 1.5 hours at 110 ° C, cooled to 0 ° C and concentrated to 200 ml. The resulting crystals were combined with 1000 ml of acetone, and the resulting precipitate was filtered off with suction, washed with acetone and dried over P4O10 (34.9 g, 47%).

Elemental analysis for C9H9N3O2S x 2HCl (296.2):

Calculated: C, 36.4; H, 3.75; N, 14.1;

S, 10.8; Q, 23.9; Found: C, 35.5; H, 4.1. N: 13.5.

S, 10.9. Cl, 23.5%.

M.p .: 190-191 ° C [j ^Z0 '589- -84.4 ° (c = 1, in HCl solution)

NMR (θ2θ): δ-5.25 (s, 1H), 7.58 (t, 2H), 7.88 (s, 1H) ppm.

Concentration of the mother liquor and digestion of the crystals formed with acetone afforded another product of the same purity (20 g, 27%).

Λ = 589--78.7 ′ (c-1, IN hydrochloric acid).

g) Preparation of Da- (tert-Butoxycarbonylamino) -α- (2-aminobenzothiazol-6-yl) acetic acid (2g) g (0.0675 mol) (2f) 310 10 ml of 10% lithium hydroxide solution and 210 ml of dioxane are mixed with 36.3 g (0.166 mol, 2.45 equivalents) of di-tert-butyl dicarbonate and stirred overnight at pH 10.

The precipitated lithium carbonate is filtered off with suction and washed with warm water (300 mL)

The filtrate was concentrated to 150 ml, acidified to pH 3.5 with 2N hydrochloric acid and extracted twice with ethyl acetate / tetrahydrofuran (10: 1).

The combined extracts were washed with brine, dried over magnesium sulfate, concentrated to a volume of 100 mL, and mixed with n-hexane (500 mL).

The precipitated product is filtered off with suction, washed with n-hexane and dried over potassium hydroxide in vacuo to yield 15.9 g.

Elemental analysis for C14H17N3O4S x H2O (341.3):

H, 5.6; N, 12.3; S, 9.4. Found: C, 48.2; H, 5.3; N, 11.3; S, 10 0%.

la] 589-130.9 '(c-1, CH 3 OH)

NMR (DMSO): δ- 1.4 (s, 9H), 5.1 (d, 1H), 7.257.34 (dd, 2H), 7.53 (d, 1H), 7.71 (s, 1H) ), 7.98 (s, 2H) ppm.

h) Preparation of D-7- (2-Aminobenzothiazole-6-U-glycylamido) 3-chloro-3-cephem-4-carboxylic acid

15.5 g (0.0479 mol) of 2g in analogy to Example 1f in 220 ml dimethylformamide with 6.78 ml (0.0479 mol) of triethylamine and 5.89 ml (0.0479) mole) of pivaloyl chloride.

To the resulting mixed anhydride at -10 ° C was added dropwise a solution of 7-ACCA triethylammonium salt in 15 ml of a mixture of 11.24 g (0.0479 mol) of 7-ACCA in 110 ml of tetrahydrofuran and 110 ml of water. by rubbing and adding dropwise triethylamine.

Simultaneously with the addition of the mixed anhydride, the reaction mixture was maintained at pH 7.5 with triethylamine in zinc / tetrahydrofuran (1: 1).

Isolation and deprotection of the Boc-protected cephalosporin was carried out analogously to Example 1f. Yield: 13.4 g (51%, crude product).

C16H14CIN5O4S2 x FC3COOH (553.9)

The crude product was chromatographed on a Whatman column (Magnum M40, 500 x 40, Partisil ODS-3, 50 ppm) using 2% acetic acid as the eluent to give 4.3 g of product.

Analysis calculated for C 16 H 14 ClN 5 O 4 S 2 x 2H 2 O x 1 / 3CH 3 COOH (495.9): C, 40.36; H, 3.93; N, 14.12.

S '12 9% Found: C: 40.5, H: 3.9, N: 12.9,

S, 12.0%.

NMR (DCOOD): δ-3.58 (d, J-18Hz, 1H), 3.94 (d, J-18Hz, 1H), 5.32 (d, J-5Hz, 1H), 5.75 (s, 1H), 5.94 (d, J-5Hz, 1H), 7.84 (s, 2H), 8.12 (s, 1H) ppm.

-9HU 200183Β

Example 3

Preparation of DL-7- (2-Amino-benzothiazol-6-yl-glycyl-amido) -3-methyl-3-cephem-4-carboxylic acid (1-2)

To a solution of 4g (1.2 mmol) of letoek (mono-Boc-5 derivative) in a mixture of 30 ml of dimethylformamide and 30 ml of tetrahydrofuran, cooled successively to 2.16. of ethyl diisopropylamine (0.4 mL, 12.4 mmol) and methanesulfonyl chloride (0.96 mL, 12.4 mmol).

The reaction mixture was stirred for 45 minutes at -50 ° C and 3.35 g (12.4 mmol) of 7-amino-3-methyl-3-cephem-4-carboxylic acid tert-butyl ester and 2.16 mL (12.4 mmol) were added dropwise. a solution of ethyl diisopropylamine in a mixture of 25 ml of tetrahydrofuran and 25 ml of methylene chloride (0 ° C). The resulting reaction mixture was stirred for 45-60 minutes. At 'C without cooling. The reaction mixture was evaporated in vacuo and the residue was dissolved in ethyl acetate (300 mL).

Wash with 0.1 N hydrochloric acid, brine, sodium bicarbonate and water. After drying and evaporation of the solvent, 3.2 g (45%) of product are obtained. 25

Boc-protected cephalosporin (1.5 g, 2.6 mmol) was dissolved in methylene chloride (30 ml), anisole (0.5 ml) and trifluoroacetic acid (30 ml) were added at 0 ° C and the reaction mixture was stirred at room temperature for 1 hour. 30

From the resulting reaction mixture, the trifluoroacetic acid / methylene chloride mixture was evaporated in vacuo and the resulting oily substance was mixed with ether. The trifluoroacetate was filtered off with suction, washed with ether, dried, taken up in 200 ml of water and applied to an Amberliter IRA-35 (acetate-form) column. The column was washed with water (200 mL) and the eluate lyophilized. Yield: 764 mg.

C17H17N5O4S2 (419.5)

NMR (DCOOD): δ 2.16 (d, 3H), 3.2-3.64 (mm. 40H), 5.18-5.26 (dd, 1H), 5.72-5.85 (m and d, 2H),

7.32 (m, 2H), 8.16 (d, 1H) ppm.

áujélda

Preparation of D-7- (2-Aminobenzothiazol-6-yl-glycylamido) -3-45 methyl-3-cephem-4-carboxylic acid and its L-form

400 mg of the product of Example 3 on a preparative Zorbax column (DuPont 250-21, ODS, 8 µm, 230 nm, eluant 93% 0.1% aqueous trifluoroacetic acid 50 solution / 7% methanol) were added to D and L. -We separate it into shape.

L-shape (vertex I):

After elution, the first peak is the L-form. Yield: 310 mg. 55

Trifluoroacetate was dissolved in water (50 mL), applied to an Amberlite 1RA-68 (acetate form) column, eluted with water (200 mL) and lyophilized. Yield: 55 mg.

C17H17N5O4S2 (419.5) 60

NMR (DCOOD): δ-2.26 (s, 3H), 3.48 (d, J-18Hz,

1H). 3.62 (d, J -18Hz, 1H), 5.26 (d, J-5Hz, 1H), 5.78 (5.III). 5.82 (<1, J-5Hz, 1H), 7.84 (q, 2H), 8.18 (s, 1H) ppm.

Form D (11 peaks): 65

Further elution with 0.1% trifluoroacetic acid / methanol gave D-peak as titanium II. Yield: 71 mg.

Trifluoroacetate was converted to betaine on an Amberlite IRA-68 (acetate) column. Yield:

47.3 mg.

Nmr (DCOOD): δ 2.19 (s, 3H), 3.3 (d J-18Hz, 1H). 3.5 (d, J-18Hz, 1H), 5.2 (d, J-5Hz, 1H), 5.75 (s 1H), 5.86 (d, J-5Hz, 1H) .7.84 ( s, 2H), 8.16 (s, 1H) ppm.

Example 5

Preparation of DL-7- (2-Amino-6-methyl-benzothiazol-4-yl-glycyllamido) -3-chloro-3-cephem-4-carboxylic acid (1-3)

a) Preparation of 2-Amino-6-methyl-benzothiazole carboxylic acid 4-methyl ester (5a) Methyl 2-amino-5-methyl-benzoic acid methyl ester (0.303 mol) was dissolved in glacial acetic acid (300 ml) and 120.6 g (1.24 mol) of potassium rhodanide in 200 ml of glacial acetic acid. Bromine (17.42 mL, 0.335 mol) was added dropwise to the resulting reaction mixture at 10 ° C and stirred for 2 hours without cooling. The resulting suspension is poured into 3 L of ice water with stirring, the precipitated product is filtered off with suction and washed with dilute sodium carbonate solution and water. The filter cake was slurried in water and stirred at 100 ° C overnight. After cooling, the product is filtered off with suction, washed with water and dried in vacuo at 40 ° C. Yield: 60g (89%).

C10H10N2O2S (222.2)

NMR (DMSO): δ-2.35 (s, 3H). 3.8 (s, 3H), $ 1 (s, 1H), 7.71 (s, 1H), 7.81 (s, 2H) ppm.

b) Preparation of 2-Amino-4- (hydroxymethyl) -6-methylbenzothiazole (5b) g (0.135 mol) of (5a) in analogy to Example 1a (337.7 mL, 0.405). mole) was reduced with DIBAL (20% toluene solution). Yield: 21.2 g (81%).

H9C10N2OS (194.2)

NMR (DMSO): δ - 2.35 (s, 3H), 4.72 (d, 1H), 5.05 (t, 2H), 7.12 (s, 1H), 7.34 (s, 1H) ), 7.41 (s, 2H) ppm.

c) Preparation of 2-Amino-methyl-benzothiazole-4-carboxaldehyde (5c)

Compound 5b (67.6 g, 0.348 mol) was treated with manganese (TV) oxide (241 g, 2.77 mol) in analogy to Example 1b. Yield: 33.6 g (50%).

C9H8N2OS (192.2)

NMR (DMSO): δ - 2.39 (s, 3H), 7.45 (s, 1H), 7.79 (s, 1H), 8.01 (s, 2H), 10.6 (s, 1H) ) ppm.

d) Preparation of 5- (2-Amino-6-methyl-benzothiazol-4-yl) -2,4-imidazolidinedione (5d)

Compound 5c (36.2 g, 0.188 mol) was reacted with ammonium carbonate (75.9 g, 0.791 mol) and sodium cyanide (14.3 g, 0.292 mol) in analogy to Example 1c. Yield: 25.5 g (52%).

Elemental analysis calculated for C11H toN402S (262.3):

Found: C, 50.37; H, 3.84; N, 21.36; S, 12.22. Found: C, 51.2; H, 5.0; N, 21.0; S, 11 , 2%.

NMR (DCOOD): δ - 2.28 (s. 3H), 5.87 (s, 1H). 7.43 (s, 1H), 7.48 (s, 1H), 7.64 (s, 2H) ppm.

-10HU 200183Β

e) Preparation of DL-α-Amino-α- (2-amino-6-methylbenzothiazol-4-yl) acetic acid (5e)

5d (25.5 g, 97.2 mol) in hydrogen bromide (48%, 500 mL) was refluxed for 24 hours. The reaction mixture is evaporated to dryness, the residue is taken up in water, washed twice with ethyl acetate and the aqueous phase is evaporated to dryness in vacuo. Yield:

38.1 g.

Elemental Analysis for: C 10 H 11 N 3 O 2 S · 2HBr · 2H 2 O (435.1) Calculated: C, 27.60; H, 3.94; N, 9.65.

S, 7.36; Br, 36.73. Found: C, 26.9; H, 3.9; N, 10.2;

S, 6.5; Br: 40.8%.

f) Preparation of DL-α- (tert-Butyloxy) carbonylamino-α- (2-amino-6-methylbenzothiazol-4-yl) acetic acid (5f)

Compound (5e) (34.7 g, 0.09 mol) was stirred with ditert-butyl dicarbonate (98.2 g, 0.45 mol) for 4 days at room temperature in analogy to Example le. Crude yield: 19.8 g (50%), after recrystallization from methanol, 6 g are obtained.

C1SH19N3O4S (337.4)

NMR (DMSO): δ-1.37 (s, 3H), 5.57 (d, 1H), 7.0 (s, 1H), 7.25 (d, 1H), 7.35 (s, 1H), 7.52 (s, 2H) ppm.

g) Preparation of DL-7- (2-amino-6-methyl-benzothiazol-4-yl-glycyl-amido) -3-chloro-3-cephem-4-carboxylic acid

3.37 g (0.01 mol) of 5f in analogy to Example 1f) were treated with 1.4 ml (0.01 mol) of triethylamine, 1.23 ml (0.01 mol) of pivaloyl chloride and Reaction with 2.34 g (0.01 mol) of 7-ACCA.

The Boc-protected vephalosporin is deprotected in an analogous manner to Example lg). To remove trifluoroacetic acid, the product is treated with Amberlite IRA-68 (acetate form) and lyophilized. Yield: 830 mg (15%).

C 17 H ₁₆ ClN ₃ O 4 S ₂ x 3H ₂ O (507.9)

NMR (DCOOD): δ - 2.49 (d, 3H), 3.58-4.02 (mm, 2H), 5.36 (dd, 1H), 5.83-5.94 (m, 2H) , 7.56 (m, 1H), 7.84 (m, 1H) ppm.

(Lpélda

Preparation of DL-6- (2-Aminobenzothiazol-6-yl-glycylamido) penicillanic acid (1-5)

a) Preparation of DL-α- (Benzyloxy) carbonylamino-α- (2-aminobenzothiazol-6-yl) acetic acid (6a) g (16.9 mmol) The dihydrochloride was suspended in 100 ml of water and heated to 100 ° C with 4.25 g (50.7 mmol) of sodium bicarbonate. To the resulting reaction mixture was added 2N sodium hydroxide solution until pH 9 was reached. To the resulting mixture was added 4.8 g (17.7 mmol) of benzyl p -nitrophenyl carbonate dissolved in 100 mL of dioxane and the mixture was stirred for 3 hours at 100 ° C.

The resulting solution was filtered hot, the dioxane removed from the filtrate in vacuo and washed twice with ethyl acetate. The aqueous phase was acidified to pH 3.8 with 2 µl hydrochloric acid at 10 ° C, filtered off with suction and dried at 40 ° C. then digested in hot acetone and mixed with ether at 20 ° C and filtered off with suction. Yield: 3 g (50%).

C17H15N3O4S (357.4)

NMR (DMSO): δ-5.07 (s, 2H), 5.18 (d, J-7.5 Hz, 1H), 7.36 (m, 7H), 7.73 (s, 1H), 7.96 (s, 2H), 8.08 (d, 1H) ppm.

b) Preparation of allyl DL-6 [α- (benzyloxy) carbonylamino-α (2-aminobenzothiazol-6-yl-glycylamido)] penicillinate (6b) g. (6 mmol) of a solution of (6a) in THF (32 mL) cooled to -50 ° C was successively sprayed with 0.98 mL (5.6 mmol) of ethyl diisopropylamine and 0.44 mL (5.6 mmol) methanesulfonyl chloride). The reaction mixture was stirred for 45 minutes at -50 ° C and added dropwise to 2.52 g (5.88 mmol) of allyl-6-aminopenylcillinate-p-toluenesulfonic acid salt and 1.02 mL (5.88 mmol) of ethyl diisopropyl. amine in 32 ml of tetrahydrofuran. The resulting reaction mixture was stirred for 15 minutes at -50 ° C and then for 45-60 minutes without cooling. The reaction mixture was evaporated in vacuo and the residue was taken up in 300 ml of ethyl acetate, 0.1 N hydrochloric acid, brine, sodium bicarbonate and water. After drying and evaporation of the solvent, 2.2 g (66%) of (4b) are obtained.

C28H29N5O6S2 (595.7)

NMR (DMSO): δ-1.32-1.60 (mm, 6H), 4.4 (d, 1H), 4.67 (d, 2H), 5.05 (d, 2H), 5.3 (dd, 1H), 5.39-5.57 (mm, 4H), 5.95 (mm, 1H), 7.3 (m, 7H), 7.5 (d, 2H), 7.7 ( d, 1H), 7.98 (m, 1H), 9.03 (q, 1H) ppm.

c) Preparation of DL-6 - [α- (Benzyloxy) carbonylamino-α- (2-amino-benzothiazol-yl] -glycylamido)] - penicillanic acid (6c)

To a solution of 6b (2.1 g, 3.53 mmol) in methylene chloride (12 mL) was sprayed with sodium caprylate (10.6 mL, 5.3 mmol) in ethyl acetate (10.6 mL, 5.3 mmol) and 92.6 mg (0.353 mmol, 0.1 equivalent) of triphenylphosphine are added. After 2 minutes, tetrakis (triphenylphosphine) palladium (0) (93.6 mg, 0.081 mmol, 2.3 mole% of compound (2b)) was added and the mixture crystallized for 20 minutes, during which time the sodium salt crystallized. . The product obtained is mixed with acetone, filtered off with suction, washed with acetone and ether. Yield: 1.8 g (87%).

C25H24NaN5O2S2 (577.6)

NMR (DMSO): δ-1.42-1.6 (q, 6H), 3.88 (d, 1H), 5.08 (s, 2H), 5.32 (d, 1H), 5.42 (d, 1H), 5.54 (d, 1H),

7.4 (m, 7H), 7.57 (s, 2H), 7.75 (s, 1H) ppm.

d) Preparation of Sodium D1-6- (2-Aminobenzothiazol-6-ylglycylamido) penicillate (6d)

5.4 g (9.35 mmol) of 6c is dissolved in 15 ml of n-butanol in 200 ml of water and hydrogenolytically deacetylated in a pre-hydrogenated aqueous solution over 30 g of palladium on carbon for 60 minutes. After separation of the catalyst and distillation of the n-butanol, the aqueous filtrate is lyophilized. Yield:

3.4 g (83%).

Ci7Hi8NaN ₅ O ₄ S2 (443.5)

NMR (DMSO): δ-1.47-1.62 (q, 6H), 3.97 (d, 1H). 5.0 (br d, 1H), 5.4 (d, 1H), 5.48 (d, 1H), 7.28 (s, 2H), 7.51 (s, 2H), 7.68 ( s, 1H) ppm.

-11HU 200183Β

Example 7

Preparation of D-6- (2-Aminobenzothiazolyl-yl-glycyl-amido) -pe nicillanic acid

3.4 g of compound (6) are separated into D and L forms on a preparative Zorbax column (DuPont 250-21, ODS, 8 μχη, 240 nm). 9 g of sodium dihydrogen phosphate in 1000 ml of water / methanol (86:14) were used as mobile phase. Methanol was distilled off from the eluate (Peak II) and the resulting solution was desalted on a Lobar column (size B, RP-8) vis /. and first elute with water. Subsequently, the column was inverted at 180 ', rinsed with acetonitrile and the eluate evaporated to dryness in vacuo to yield 120 mg.

NMR (DMSO): δ-1.38 (s, 3H), 1.48 (s, 3H), 4.22 (s, 1H), 5.01 (s, 1H). 5.31 (d. J-5Hz, 1H). 5.4 (d, J-5Hz, 1H), 7.3 (m, 2H), 7.64 (s, 2H), 7.72 (s, 1H) ppm.

Example 8

Preparation of DL-7- (Benzimidazol-5 (6) -ylglycylamido) -3-chloro-3-cephem-4-carboxylic acid (1-4)

a) Preparation of 5 (6) - (hydroxymethyl) -benzimidazole (8a): Methyl ester of benzimidazole carboxylic acid 5 (6) (0.227 mole) in tetrahydrofuran at 0 ° C in analogy to Example 1a) 1150.3 mL (1.38 mol) of DIBAL (20% solution in toluene, Schering) was stirred overnight at room temperature (22.5 g, 67%).

C8H ₈ N2O (148.2)

NMR (DMSO): 8- 4.61 (s, 2H), 5.22 (broad, 1H), 7.18 (d, 1H), 7.56 (d, 2H), 8.2 (s, 1H ), 12.42 (s, 1H) ppm.

b) Preparation of benzimidazole-5 (6) -carboxaldehyde (8b)

Compound (8a) (42.3 g, 0.285 mol) in tetrahydrofuran (130 ml) and dimethylformamide (1300 ml) was stirred with manganese (TV) oxide (175 g, 2.01 mol) at room temperature for 48 hours and then manganese (IV) on a Seitz filter. oxide was separated and the filtrate was evaporated to dryness. The residue was stirred in petroleum ether, filtered off with suction and dried in vacuo at 60 ° C (37.3 g, 89%).

C ₈ H ₆ N ₂ O (146.1).

NMR (DMSO): δ- 7.78 (d, 2H), 8.22 (s, 1H), 8.42 (s, 1H), 10.05 (s, 1H) ppm.

c) Preparation of 5- (Benzimidazol-5- (6) -yl) -2,4-imidazolidinedione (Compound 8c)

Compound 8b (45.8 g, 0.313 mole) was treated with 23.03 g (0.47 mole) of sodium cyanide and 120.3 g (60%) in ethanol / water at 60 ° C for 20 hours. 1.25 mol) of ammonium carbonate. Yield: 36.4 g (54%).

C10H8N4O2 (216.2)

NMR (DMSO): δ-5.44 (s, 1H), 7.53 (d, J-9Hz, 1H). 7.83 (s, 1H), 7.88 (d, J-9Hz, 1H), 8.63 (s, 1H). 9.63 (s 1H). 10.93 (s, 1H) ppm.

d) Preparation of DL-α-Amino-α- (benzimidazol-5 (6) -yl) acetic acid 7DL-benzimidazol-5 (6) -ylglycine), compound (8d) g (0.093 mol) (8c) and 105.7 g (0.335 mole) (3.6 equivalents) of barium hydroxide in 1000 ml of water were stirred at 100 ° C for 24 hours. The resulting slurry was diluted with 500 mL of water at 100 ° C, bubbled with carbon dioxide gas for 2 hours, filtered off with suction, and washed with hot water. The filtrate was evaporated to dryness. Yield: 20.1 g (80%).

Analysis for C9H9N3O2 x 2H ₂ O xl / 3BaCO3 (293.0) Calc'd: C: 38.26, H: 3.10, N: 14.34,

Ba: 15.62 Found: C: 40.6, H: 3.9, N: 15.5.

Ba: 14.7%.

NMR (DCOOD): δ-5.71 (s, 1H), 7.94 (d, 1H). 8.16 (d, 1H), 8.86 (s, 1H), 9.5 (s, 1H) ppm.

e) DL-α- (tert-Butyloxy) carbonylamino-α- (1-tert-butyl-oxy-carbonyl-benzimidazole-5 (6) -fl) -acetic acid / (8e) (844 g) (0.044 mole) was stirred with di-tert-butyl dicarbonate (38.4 g, 0.176 mole) overnight at room temperature in analogy to Example le. The crude product was dissolved in ethyl acetate (80 ml) and stirred at room temperature for 6 hours. ml of light petroleum. Yield: 14.6 g (84%).

Elemental analysis Ci9H sN3O6 ₂ (391.4) Calc'd:

Found: C, 58.30; H, 6.44; N, 10.73. Found: C, 56.80; H, 6.60; N, 9.80.

NMR (NaOD): δ - 1.24 (s, 9H), 1.4 (s, 9H), 5.05 (s, 1H), 7.2 (d, 1H), 7.65 (s and d) , 2H), 8.06 (s, 1H) ppm.

d) Preparation of DL-7- (Benzimidazol-5 (6) -yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid x trifluoroacetate (8f)

Compound 8e (2.0 g, 5.1 mmol) in analogy to Example 1f was treated with triethylamine (0.71 mL, 5.1 mmol), pivaloyl chloride (0.628 mL, 0.51 mol) and 32 g (5.61 mmol) of 7-ACCA are reacted with 2.3 g (3.78 mmol) of Boc-protected cephalosporin in a manner similar to that described in Example lg). The trifluoroacetate was dissolved in water, washed twice with ethyl acetate / ether (1: 1) and the aqueous phase was lyophilized. Yield: 1.5 g (76%).

C 16 H 14 ClN 5 O 4 S x CF 3 COOH (521.9)

NMR (DCOOD): δ- 3.4-4.03 (mm, 2H), 5.29-

5.38 (dd, 1H), 5.88-5.96 (dd and s, 2H), 7.93-8.0 (t, 1H), 8.16-8.22 (t, 1H), 8.3 (s, 1H); 9.52 (m, 1H).

Example 9

Preparation of D-7- (Benzimidazol-5 (6) -yl-glycylamido) -3-chloro-3- (cephem-4-carboxylic acid) and its L-form

700 mg of compound (8f) is separated on D and Lala by preparative HPLC (Hibar 250-25, RP-18, 7 µm, 220 nm, running 1000 µl water / 5 ml acetonitrile / 1 ml acetic acid), Portions (30 mg) of DL (8f) were dissolved in 2 mL of runner system, applied to the column and separated into 2 fractions (peak I and peak II) at runner speed of 10 mL / min.

(a) Form D (peak II):

Yield: 18 mg

HPLC analytical determination: identical to that given for L-form.

Retention 8.14

-12HU 200183Β

(b) L-shape (vertex I):

Yield: 42 mg

HPLC analytical determination: (Hibar 250-4, RP- 8, 10 µm, 255 nm, mobile phase: 975 ml of pH 7 Merck phosphate buffer, 25 ml of acetonitrile, flow rate:

1.5 ml / min).

Retention: 6.90

NMR (DCOOD): 8- 3.74 (d, J-18Hz, 1H), 3.98 (d, J-18.5Hz, 1H), 5.35 (d, J-5Hz, 1H), δ, 86 (s, and d, 2H). 7.96 (d, 1H). 8.16 (d, 1H), 9.5 (s, 1H) ppm.

Example 10

Preparation of DL-7- (Benzimidazol-5 (6) -yl-glycyl-amido) -3-methyl-3-cephem-4-carboxylic acid (I-6)

3.5 g (8.9 mmol) of 8e and 1.36 g (8.9 mmol) of 1-hydroxybenzotriazole are dissolved in 15 mL of tetrahydrofuran under a nitrogen atmosphere. To the solution was added 1.84 g (8.9 mmol) of N, N'-dicyclohexylcarbodiimide (DCC) in 10 mL of tetrahydrofuran and stirred at room temperature for 2 hours. (9 mmol) of 7 amino-3-methyl-3-acephem-4-carboxylic acid tert-butyl and the resulting mixture was stirred overnight without cooling. The precipitated urea is filtered off with suction, washed with tetrahydrofuran and the filtrate is evaporated to dryness. The residue was dissolved in ethyl acetate, washed with sodium bicarbonate solution / water, dried over sodium sulfate and the filtrate was evaporated to dryness in vacuo (3.7 g). Chromatography on a silica gel column with petroleum ether / ethyl acetate (3: 1) and petroleum ether / ethyl acetate (1: 1) gave 1.3 g (23%) of pure product.

1.0 g (1.55 mmol) of Boc-protected cephalosporin is deprotected in analogy to Example 3, the trifluoroacetic acid is removed on an Amberlite IRA-68 (acetate form) column and the resulting solution is lyophilized. Yield: 260 mg (43%).

C 17 H 17 N 5 O 4 S (387.4)

NMR (DCOOD): 8-2.18 (s, 3H), 3.2-3.65 (mm, 2H), 5.18-5.26 (dd, 1H), 5.78-5.88 ( dd, 2H), 7.98 (d, 1H), 8.18 (t, 1H), 9.5 (d, 1H) ppm.

JJLpéldA

Preparation of D1-7- (2-Amino-1H-benzimidazol-5 (6) -yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid (1-7)

a) Preparation of 2-Amino-5 (6) -hydroxymethylbenzimidazole (IIa)

132 g (0.745 mol) of methyl 2-amino-5 (6) -benzimidazole carboxylate are suspended in 2000 ml of tetrabydrofuran under a nitrogen atmosphere. To the resulting slurry, 596 ml (2.08 mol) of sodium bis (2-methoxyethoxy) aluminum hydride (Red-Al 3.5 mol) was added dropwise over 2 hours under gentle cooling (ice bath) at 25-31 ° C. toluene solution) in THF (1000 mL). The resulting solution was then stirred for 1 hour without cooling and refluxed overnight. Excess Red-Al was carefully quenched by the addition of 160 mL of water under nitrogen, the resulting reaction mixture was stirred for one hour, the aluminum hydroxide was suctioned off, washed with 2 L of tetrahydrofuran and the filtrate was evaporated to a dark oil. The residue was taken up in 500 mL of ethyl acetate and washed three times with 500 mL of water. The combined aqueous layer was washed with ethyl acetate (3 x 200 mL), the aqueous layer was filtered and lyophilized. Yield: 74 g (61%).

C8H9N3O (163.2)

NMR (DMSO): 8- 4.46 (s, 2H), 6.24 (s, 2H), 6.82 (d, 1H), 7.03 (d, 1H), 7.09 (s, 1H) ) ppm.

b) Preparation of 2-Aminobenzimidazole-5 (6) -carboxaldehyde (11b) Compound (11a) (0.453 mole) in glacial acetic acid (200 ml) at room temperature for 3 days at 248.4 g (2.857 mole). (3 times excess) with manganese dioxide. The manganese dioxide is filtered off with suction, the filtrate is again filtered through silica gel and the resulting acetic acid solution is evaporated. The residual oil was digested with ethyl acetate (1.5 L), whereupon crystallization began. The resulting crystalline material is filtered off with suction, washed with ether and dried in vacuo.

Yield: 119.7 g (96%).

H, 6.22; N, 15.26. Found: C, 44.20; H, 4.90; C ,HH 44,5NNO x. N, 11.00%.

c) Preparation of 5- (2-Amino-1H-benzimidazol-5-yl) -2,4-imidazolidinedione (11c)

119.7 g (0.435 mole) of 11b are mixed with 219 g (2.28 mole) of ammonium carbonate and 41.4 g (0.845 mole) of sodium cyanide in 900 ml of ethanol in a manner similar to that described in Example 1c. at 900 ° C for 20 hours in 900 ml of water. After distilling off the ethanol, the resulting slurry is acidified at 0 ° C to pH 2 and then adjusted to pH 6.0 to give an oily product which slowly solidifies. Yield: 57.6 g (58%).

Elemental analysis for C10H8N5O2 (231.2):

Found: C, 51.94; H, 3.92; N, 30.28. Found: C, 51.40; H, 4.10; N, 29.10.

d) DL-α- (tert-butyloxy) carbonylamino-α- (2-tert-butyloxycarbonyl-1H-benzimidazol-5-yl) acetic acid (1d) / production

Compound 11c (41.2 g, 0.178 mol) was stirred with lithium hydroxide (42.6 g, 1.781 mol) in water (1000 mL) at 100 ° C for 2 days in analogy to Example 1d. The resulting solution was filtered hot, the filtrate was acidified to pH 2 at 0 ° C and the aqueous phase was extracted with ethyl acetate. The aqueous solution was evaporated to dryness in vacuo to yield 62.4%.

NMR (NaOD, DMSO): 8-4.12 (s, 1H), 6.71 (d, 1H), 6.97 (d, 1H), 7.01 (s, 1H) ppm.

The residue was converted into a solution of sodium hydrogencarbonate (25.4 g, 0.303 mol) in water (300 ml) and di-tert-butyl dicarbonate (66.1 g, 0.303 mol) was added dropwise. The reaction mixture was stirred for 2 days at room temperature. The dioxane was distilled off, the residue was diluted with water and washed with petroleum ether. The aqueous phase was acidified to pH 2.5 with 2N hydrochloric acid while cooling with ice and extracted with ethyl acetate. Brine

After washing with Na2SO4, drying over sodium sulfate and evaporation of the organic phase, 17.9 g of crude product are obtained, which is precipitated from tetrahydrofuran / petroleum ether. Yield: 7.0 g.

C19H26N4O6 (406.4)

NMR (NaOD): δ-1.42 (s, 9H), 1.46 (s, 9H), 6.98 (d, 1H), 7.28 (d, 2H) ppm.

e) Preparation of D1 -7- (2-Amino-1H-benzimidazol-5-yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid (1e)

4.9 g (2.2 mmol) of compound (11d) in analogy to Example 1f) with triethylamine (1.9 ml, 12.1 mmol) and pivaloyl chloride (1.49 ml, 12.1 mmol) and Reaction with 98 g (12.7 mmol) of 7-ACCA. Yield: 3.0 g (40%; tetrahydrofuran / petroleum ether).

Boc-protected cephalosporin was treated with trifluoroacetic acid / methylene chloride (1: 1) in analogy to Example 1g.

2.35 g (91%).

C 16 H 15 ClN 6 O 4 S x CF 3 COOH (536.9).

NMR (DCOOD): δ - 3.52 ^ 4.0 (mm, 2H), 5.28-

5.36 (dd, 1H), 5.70 (d, 1H), 5.84-5.92 (dd, 1H), 7.58-7.66 (m, 2H), 7.76 (d, 1H) ppm.

Example 12

Preparation of D-7- (2-Amino-1H-benzimidazol-5-yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid and its L-form

Preparative HPLC separation of (IIe):

Column: Hibar 250-25 (RP-18.7 pjn, 254 nm)

The starting material was 1000 ml water / 60 ml acetonitrile / 1 ml trifluoroacetic acid

Method of Application: 2.3 g; always 200 mg is dissolved in 2-3 ml of running agent per column flow.

Yield: peak I (L-form): 430 mg, Π peak (Dalak): 220 mg.

A solution of 220 mg of peak II was dissolved in 15 ml of water on an Amberlite IRA-68 (acetate-form) column and washed with 200 ml of water.

The filtrate was filtered through a syringe through a Milllex-GS (0.22 µm) filter and lyophilized. Yield: 105 mg.

NMR (DCOOD): δ- 3.54 (d, J-18HZ, 1H), 3.86 (d, J-18H7, 1H), 5.28 (d, J-5Hz, 1H), 5.68 ( s, 1H), 5.92 (d, J-5Hz, 1H), 7.58 (d, 1H), 7.66 (d, 1H), 7.76 (s, 1H) ppm.

Example 13

Preparation of DL-7- (2-Amino-1H-benzoimidazol-5-yl-glycyl-amino) -3-methyl-3-cephem-4-carboxylic acid (compound of formula I-8)

Compound (1d) (4.0 g, 9.84 mmol) was treated with ethyl diisopropylamine (1.71 mL, 9.84 mmol) in methane (0.76 mL, 9.84 mmol) in analogy to Example 3. sulfonyl chloride and 2.7 g (9.84 mmol) of methanesulfonyl chloride and 2.7 g (9.84 mmol) of 7-amino-3-methyl-3-cephem-4-carboxylic acid tert-butyl ester. The product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 1: 1) to give 0.9 g.

0.8 g of Boc-protected cephalosporin was treated with trifluoroacetic acid and then with Amberlite IRA-68 (acetate-like) as described in Example 3. Yield: 420 mg.

Analysis calculated for C17 H18 N6 O4 S x 2 1/2 H2O (447.5): C, 45.63; H, 5.18; N, 18.78.

S, 7.16. Found: C, 45.50; H, 5.00; N, 16.90.

S, 6.60%.

NMR (DCOOD): δ-2.2 (s, 3H), 3.28-3.69 (mm, 2H). 5.21-5.3 (dd, 1H), 5.75 (d, 1H), 5.82-5.89 (dd, 1H), 7.62-7.74 (m, 2H), 7, 82 (s, 1H) ppm.

Example 14

Preparation of DL-7- (2- (Trifluoromethyl) -1H-benzimidazole-5-flglycylamido) -3-chloro-3-cephem-4-carboxylic acid (1-9)

a) Preparation of 2- (Trifluoromethyl) -1H-benzimidazole-5-carboxylic acid (14a): 3,4-Diaminobenzoic acid (g, 0.329 mol) in 4N HCl (300 mL) 40.5 g (0.355 mol) of trifluoroacetic acid was refluxed for 5 hours. The resulting solution was cooled to 0 ° C, the precipitated product was filtered off with suction and dried in vacuo at 40 ° C (59.7 g, 73%).

Elemental analysis based on C9H5F3N2O2 x H2O (248.2):

H, 2.84; N, 11.28 Found: C, 4336;

F, 22.98; Found: C, 42.60; H, 2.80; N, 10.90.

F: 22.00%.

b) Preparation of 2- (trifluoromethyl) -1H-benzimidazole carboxylic acid 5-methyl ester (14b)

A solution of (14a) (59.7 g, 0.241 mol) in methanol (1000 mL) was bubbled through hydrogen chloride for 2 hours while being heated to reflux. From the resulting reaction mixture, methanol was distilled off and the residue was dissolved in ethyl acetate / water and adjusted to pH 7 with 2N sodium hydroxide. The ethyl acetate layer was separated, the aqueous layer was extracted twice with ethyl acetate, the organic layer was washed with brine, dried over sodium sulfate and evaporated. Yield: 54.2 g (92%).

Elemental analysis for C10H7N2O2F3 (244.2):

Calculated: C, 49.19; H, 2.89; N, 11.47; F, 23.34. Found: C, 49.60; H, 3.10; N, 11.30; F, 22.40.

NMR (DMSO): δ- 4.9 (s, 3H), 7.82 (d, 1H), 8.0 (dd, 1H), 8.30 (s, 1H) ppm.

c) Preparation of 2- (trifluoromethyl) -5- (hydroxymethyl) -1H-benzimidazole (14c)

Compound 14b (53.4 g, 0.219 mol) was stirred with DIBAL (20% solution in toluene) (728.9 ml, 0.875 mol) in 600 ml of tetrahydrofuran in analogy to Example 1a) overnight at -70 ° C. . Crude yield: 45.4 g.

Elemental analysis for C9H7F3N2O (216.2):

H, 3.26; N, 12.96 Found: C, 50.0;

F: 26.37; Found: C: 49.70. H: 3.70, N: 11.70,

F: 26.00%.

On a silica gel 60 (Merck 0.04-0.063 mm) column toluene / ethyl acetate (3: 1), toluene / ethyl acetate (1: 1), toluene14

Chromatography using a 1: 3 mixture of ethyl acetate and ethyl acetate (1: 3) as eluant gave 29.3 g (62%) of benzimidazole alcohol.

NMR (DMSO): 6- 4.64 (s, 2H), 5.32 (broad, 1H), 7.36 (d, 1H), 7.66 (s 1H), 7.71 (d, 1H) ) ppm.

d) Preparation of 2- (trifluoromethyl) -1H-benzimidazole-5-carboxaldehyde (14d)

Compound 14c (29.3 g, 0.136 mol) was stirred with 71.4 g (0.821 mol) of manganese (TV) oxide in 1000 ml of tetrahydrofuran, in analogy to Example 1b, for 48 hours. Yield: 23.8 g (82%).

C9H5F3N2O (214.2)

NMR (DMSO): δ- 7.87 (q, 2H), 8.32 (s, 1H), 10.11 (s, 1) ppm.

e) Preparation of 5- (2- (trifluoromethyl) -1H-benzimidazol-5-yl) 2,4-imidazolidinedione (14e)

Compound 14d (29.2 g, 0.136 mol) was reacted with ammonium carbonate (55 g, 0.573 mol) and sodium cyanide (10.4 g, 0.212 mol) in ethanol / water in analogy to Example 1c. After distilling off the ethanol, the solution was acidified to pH = 2 with 2N hydrochloric acid solution at 0 ° C, then the pH was restored to 4.5 and extracted several times with ethyl acetate. The combined ethyl acetate layer was washed with brine, dried and evaporated. Yield ,? g (80.6%).

C 1 H 7 F 3 N 4 O 2 (284.2).

NMR (DMSO): δ-5.85 (s, 1H), 7.34 (d, 1H), 7.69 (s, 1H), 7.77 (d, 1H), 8.5 (s, 1H), 10.83 (s, 1H) ppm.

f) Preparation of DL-α-Amino-α- (2- (trifluoromethyl) -1H-benzimidazol-5-yl) acetic acid (14f)

Compound 14e (35.7 g, 0.126 mol) was reacted with 500 ml of hydrogen bromide (48%) in analogy to Example 5e. Yield: 18.1 g (42%).

C 10 H 8 F 3 N 3 O 2 x HBr (340.0)

NMR (DCOOD): δ-5.88 (s, 1H), 8.1 (d, 1H), 8.28 (d, 1H), 8.46 (s, 1H) ppm.

g) Preparation of DL-α- (tert-Butyloxy) carbonylamino-α- (2-trifluoromethyl / -1H-benzimidazol-5-yl) -acetic acid (14g)

14f (23.6 g, 0.069 mol) was dissolved in a mixture of 10% sodium bicarbonate solution (150 mL) and dioxane (150 mL), treated with ditert-butyl dicarbonate (48.2 g, 0.221 mol) and overnight. The dioxane is distilled off, the aqueous phase is extracted with petroleum ether and acidified to pH 2 with 2N hydrochloric acid in the presence of ethyl acetate at 0 ° C. The ethyl acetate layer was separated, washed with brine, dried over sodium sulfate and evaporated.

The crude product was dissolved in ether, insoluble materials were filtered off, and the remaining solution was mixed with petroleum ether and filtered off. Yield: 10.8 g (43%).

Elemental analysis based on C15H16F3N3O4 (359.3):

H, 4.49; N, 11.69 Found: C, 50.14;

F, 15.86. Found: C, 50.10; H, 4.80; N, 12.40.

F: 15.10%.

h) Preparation of DL-7- (2- (Trifluoromethyl) -1 H -betimidazol-5-yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid (14h)

Compound 14g (4.0 g, 11.1 mmol) was treated with triethylamine (1.56 mL, 11.1 mmol), pivaloyl chloride (1.4 mL, 11.1 mmol) and 2, Reaction with 92 g (12.4 mmol) of 7-ACCA Yield: 5.1 g (89%)

The Boc-protected cephalosporin is deprotected in the same manner as in Example 1g and converted to betaine Yield: 2.0 8 (36%).

Elemental Analysis for: C 17 H 13 ClF 3 N 5 O 4 S x 1 1/2 H 2 O (502.9) Calculated: C 40.61, H 3.20, N 13.93.

S, 6.37; F, 11.34. Found: C, 40.20; H, 3.60; N, 13.60.

S, 4.80; F, 12.20%.

Example 15

Preparation of D-7- (2- (Trifluoromethyl) -1H-benzimidazol-5-yl-glycidamido) -3-chloro-3-cephem-4-carboxylic acid and its L-form

Preparation of Compound (14h) by preparative high performance liquid chromatography

Column: Hibar 250-25 (RP-18, 7 pm, 254 nm)

The starting material was 2500 ml water / 250 ml acetonitrile / 23 ml trifluoroacetic acid

Flow rate: 10 ml / min.

L-shape (vertex I):

1.4 g of compound (14h) are suspended in 20 ml of runner, leaving 400 mg of insoluble material which is the pure L-form.

NMR (DCOOD): δ- 3.7 (d, 1H), 3.93 (d, 1H), 5.3 (d, 1H), 5.78 (s and d. 2H), 7.8 (d) , 1H), 8.05 (d, 1H), 8.23 (s, 1H) ppm.

D-shape (vertex II):

The filtrate was filtered through a syringe through a Millex filter and applied to the column at 0.5 ml / min. The eluate is lyophilized. Yield: 3000 mg.

NMR (DCOOD): δ- 3.5 (d, J-18.5Hz, 1H), 3.85 (d, J-18.5Hz, 1H). 5.25 (d, J-5Hz, 1H), 5.78 (s, 1H), 5.9 (d, J-5Hz, 1H), 7.78 (d, 1H), 8.06 (d. 1H), 8.21 (s, 1H) ppm.

Example 16

Preparation of D1-6- (2- (Trifluoromethyl) -1H-benzimidazol-5-ylglycylamido) penicillanic acid (I-10)

a) Preparation of DL-α- (Benzyloxy) carbonylamino-α- (2- (trifluoromethyl) -1H-benzimidazol-5-yl) acetic acid (Compound 16a)

9.2 g (27.1 mmol) of 14f are dissolved in 50 ml of water and adjusted to pH 9 with 2N sodium hydroxide solution. The resulting clear solution was cooled to 0-5 ° C and 8.1 ml (0.0569 mol) of idoric formate benzyl ester (pH 8-10) was added dropwise over 30 minutes with the simultaneous addition of 2N sodium hydroxide solution. . The reaction mixture was stirred for 50 minutes at room temperature, extracted once with ether / ethyl acetate (1: 1), the aqueous phase was acidified to pH 2 with 2N hydrochloric acid and extracted with ethyl acetate.

After washing with sodium sulfate, the residue was precipitated from ethyl acetate / petroleum ether. Yield: 2.8 g (26%).

C18H14F3N3O4 (39.3.3)

NMR (DMSO): δ 5.09 (t, 2H), 5.38 (d, 1H), 7.34 (m, 5H), 7.47 (d, 1H), 7.73 (d, 1H) ), 7.78 (s. 1H), 8.23 (d, 1H) ppm.

b) Allyl DL 6 [((Benzyloxy) carbonylamino-a (2- (trifluoromethyl) -1H-benzimidazol-5-yl-glycyl-amido)] - penicillinate / (16b) / production

Compound 16a (2.6 g, 6.61 mmol) was treated with ethyl diisopropylamine (1.15 mL, 6.61 mmol), methanesulfonyl (0.512 mL, 6.61 mmol) in analogy to Example 6b. kforidide and a solution of 3.0 g (6.95 mmol) of p-toluenesulfonic acid salt of all-o-aminopenicillinate in 1.21 mL (6.95 mmol) of ethyl diisopropylamine in tetrahydrofuran / dimethylformamide. Yield: 3.5 g (83%).

C29H28F3N5O6S (631.6).

c) Preparation of sodium DL-6- (2- (trifluoroethyl) -1H-benzimidazole-5-yl-glycylamido) penicillinate (16c)

3.5 g (5/54 mmol) of 16b are hydrogenolytically deacylated as in Example 6d. 1.8 g (3.62 mmol) of allyl ester were treated with 10.7 ml (5.43 mmol) of 0.5 M sodium caprylate, 95 mg (0.363 mmol) of triphenylphosphine in analogy to Example 4c) and 96.2 mg (0.083 mmol) of tetrakis (triphenylphosphine) palladium (O) were treated with tetrahydrofuran under a nitrogen atmosphere.

Yield: 1.1 g (65%).

Ci8Hi7F O4SNa ₃ N ₅ (479.4).

NMR (DMSO): δ-1.43-158 (m, 6H), 3.91 (d, 1H), 453 (d, 1H), 5.33-5.44 (m, 2H), 7.05 (m, 1H),

7.38 (d, 1H), 7.5 (s, 1H) ppm.

LLfiélda

Preparation of D-6- (2- (Trifluoromethyl) -1H-benzimidazol-5-ylglycylamido) penicillanic acid and its L-form

Compound (16c) (700 mg) was pumped onto a column of a preparative Hibar column (Merck 250-25, 7 pm, 254 nm, running 2250 ml water / 125 ml acetonitrile / 125 ml acetic acid) in 10 ml of running material and D- and L- divide into shape.

L-shape (vertex I):

Yield: 110 mg

Elemental: Calculated for C 18 H 18 F 3 N 5 O 4 S × 2 H 2 O • 1/3 CH 3 COOH (513.5): C, 42.10; H, 4.58; N, 13.64. Found: C, 41.20; , 70, N: 12.10%.

NMR (DMSO): δ-1.47 (s. 3H). 158 (s. 3H), 4.14 (s, 12H), 4.9 (s, 1H), 5.48 (t, 2H), 7.47 (d. 1H), 7.74 (d. 1H) ), 7.82 (s. 1H) ppm.

HPLC: Hibar 250-4, RP-8.10 µm, 254 nm, eluent 900 ml water / 50 ml acetonitrile / 50 ml acetic acid, flow rate 3 ml / min. 1 mg / ml, retention 6.2 (product content 86.6%).

D-shape (vertex II):

Yield: 129 mg

NMR (DMSO): δ-1.38 (s, 3H), 15 (s, 3H). 4.1 (s, 1H), 4.95 (s, 1H). 5.39 (d, J-4Hz, 1H), 5.5 (s, 1H), 7.48 (d, 1H). 7.74 (d, 1H). 7.82 (s, 1H) ppm.

HPLC: identical to that obtained for L-form, retention: 9.11 (product content 93.5%).

Example 18

Preparation of DL-7- (2-Methyl-1H-benzimidazol-5-yl-glycyl-amido) -3-chloro-3-cephem-4-carboxylic acid (I-11)

a) Preparation of 2-Methyl-1H-benzimidazole-5-carboxylic acid (18a)

3,4-Diaminobenzoic acid (100 g, 0.657 mol) was refluxed with glacial acetic acid (47.3 g, 0.788 mol) and 600 ml of 4N hydrochloric acid in analogy to Example 14a). Yield: 1085 g (72%).

Elemental analysis based on C9H8N2O2 (190.2):

Calc'd: C, 63.18; H, 5.30. N, 14.73. Found: C, 63.1; H, 5.30; N, 14.60.

c) Preparation of 2-Methyl-5-hydroxymethyl-1H-benzimidazole (18c)

27.8 g (0.146 mol) of (18b) was stirred as in Example 1a) in 500 ml of tetrahydrofuran with 365.4 ml (0.438 mol) of DIBAL (20% toluene solution) overnight at -70 ° C. . The crude product obtained is stirred in hot ethyl acetate and, after cooling to 20 ° C, the residue is filtered off with suction. Yield: 10.2 g (43%).

C9H10N2O (162.2)

NMR (DMSO): δ - 2.51 (s. 3H), 455 (s. 2H). 7.06 (d, 1H), 7.38 (s, and d, 2H) ppm.

d) Preparation of 2-Methyl-1H-benzimidazole-5-carboxaldehyde (18d)

18c (24.4 g, 0.15 mol) was stirred with 84.8 g (0.975 mol) of manganese (TV) oxide in 400 ml of dimethylformamide for 3 days at room temperature.

Yield: 13.9 g (58%).

C9H8N2O (160.2)

NMR (DMSO): δ- 256 (s, 3H), 7.61 (d, 1H), 7.71 (d, 1H), 8.06 (s, 1H), 10.02 (s, 1H) ppm .

e) Preparation of DL-α-Amino-α- (2-methyl-1H-benzimidazol-5-yl) acetic acid (18e)

(18d) (22.7 g, 0.142 mol) was reacted with ammonium carbonate (57.2 g, 0596 mol) and sodium cyanide (10.8 g, 0.22 mol) in ethanol / water as described in Example le). 60 ° C for 20 hours.

After distilling off the ethanol, the solution was acidified to pH = 2 with 2N hydrochloric acid and then adjusted to pH 4.5 with 2N sodium hydroxide. The resulting clear solution was extracted twice with ethyl acetate and the aqueous phase was evaporated to dryness in vacuo. Yield: 49.0 g (containing sodium chloride).

The material was reacted with 500 ml of hydrobromic acid (48%) in analogy to Example 3c). Yield: 56.8 g.

CtoHnN ₃₀ 02x2HBr (367.1)

NMR (DCOOD): δ-5.82 (s, 1H), 7.88 (d. 1H), 8.05 (d, 1H), 8.22 (s, 1H) ppm.

f) Preparation of DL-α- (tert-Butyloxy) carbonylamino-α- (1-tert-butyloxycarbonyl-2-methylbenzimidazol-5-yl) acetic acid (18f).

-16HU 200183Β

634 g (0472 moles) of 18e & 14g are reacted with 112.6 g (0.516 moles) of di-tert-butyl dicarbonate in sodium bicarbonate solution and dioxane in an analogous manner to Yield: 12.6 g.

C20H27N3O6 (405.4).

NMR (DMSO): δ-13.5 (d, 9H), 1.65 (d, 9H), 5.22 (q, 1H), 7.36 (m, 1H), 7.65 (m, 1H) ), 7.96 (d. 1H) ppm.

g) Preparation of DL-7- (2-Methyl-1H-benzimidazol-5-yl-glycylamide) -3-chloro-3-cephem-4-carboxylic acid (18g)

4.2 g (10.3 mmol) of 18f in analogy to Example 1f) were treated with 1.44 mL (10.3 mmol) of triethylamine, 1.27 mL (10.3 mmol) of pivaloyl chloride and Reaction with 2.65 g (11.4 mmol) of 7-ACCA. Yield: 4.6 g (72%).

C27H32CIN5O8S (622.1)

4.0 g (6.43 mmol) of Boc-protected cephalosporin were deacylated in analogy to Example 1g. Yield: 2.9 g (85%).

C 17 H 16 ClN 5 O 8 S x CF 3 COOH (535.9).

NMR (DCOOD): δ-3.02 (s, 3H), 3.48-4.0 (mm, 2H), 537-5.36 (dd, 1H), 5.82 (s, 1H), 5.84 -5.94 (dd, 1H), 7.86 (d, 1H), 8.02 (d, 1H), 8.16 (s, 1H) ppm.

1.9. example

Preparation of D-7- (2-Methyl-1H-benzimidazol-5-yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid and its L-form

Preparative HPLC separation of (18g):

Column: Whatman Partisii-10,500 x 22 mm, 10 µn (Magnum 20 CCS-RP-8) 254 nm

The starting material was 2500 ml water / 25 ml trifluoroacetic acid.

Flow rate: 15 ml / min (paper speed 5 mm / min).

Method of Application: 1100 mg, 18 g in 9 ml of runner, filtered through a Millex GS, 032 µm filter.

L-shape (vertex I):

Yield: 355 mg (trifluoroacetate)

NMR (DCOOD): δ - 3.06 (s, 3H), 3.75 (d, J 8 Hz, 1H), 3.98 (d, J 8 Hz, 1H), 5.37 (d J J 5Hz, 1H), 5.84 (s, 1H), 5.88 (d, J-5Hz, 1H), (d, J-9Hz, 1H), 8.04 (d, J-9Hz, 1H), 8.18 (s, 1H) ppm.

D-shape (Π vertex):

Yield: 114 mg (trifluoroacetate)

NMR (DCOOD): δ - 3.04 (s, 3H), 3.54 (d, J18Hz, 1H), 3.87 (d, J18Hz, 1H), 539 (d, J5Hz, 1H) , 5.82 (s, 1H), 5.94 (d, J-5Hz, 1H), 7.86 (dd. 1H), 8.04 (d, 1H), 8.16 (s, 1H) ppm .

2ÍLpélda

Preparation of DL-7- (2,3-Dihydro-2-oxo-1H-benzimidazol-5-yl) glycylamido) -3-chloro-3-cephem-4-carboxylic acid (1-12)

a) Preparation of 2,3-Dihydro-2-oxo-1H-benzimidazole carboxylic acid 5-methyl ester (20a) g (0.12 mol) of 3,4-diaminobenzoic acid methyl ester and 24, 8 g (0.12 mol) of Ν, Ν'-dicarbmethoxy-Smethylisothiourea is heated to reflux in 180 ml (limethylformamide for 4 hours). After drying the product in vacuo at 40 ° C, the product was suspended in acetone and drained (21.9 g, 95%).

C9H8N2O3 (192.2)

NMR (DMSO): δ-3.8 (s, 3H), 7.01 (d, 1H), 7.48 (s, 1H), 7.64 (d, 1H), 10.9 (s 1H) ), 11.08 (s, 1H) ppm.

b) Preparation of 2,3-Dihydro-2-oxo-5- (hydroxymethyl) -1H-benzimidazole (20b)

Compound 20a (24.6 g, 0.128 mol) was suspended in tetrahydrofuran and stirred with DIBAL (20% solution in toluene) (624.8 mL, 0.768 mol) overnight at -70 ° C. Yield: 163 g (77%).

C8H8N2O2 (164.2)

NMR (DMSO): δ- 4.48 (s, 2H), 5.12 (bs, 1H), 6.91 (s, 2H), 6.98 (s, 1H), 10.61 (bs) , 2H) ppm.

c) Preparation of 2,3-Dihydro-2-oxo-1H-benzimidazole-5-carboxaldehyde (20c)

Compound 20b (25.7 g, 0.157 mol) was mixed with manganese (TV) oxide (81.7 g, 0.939 mol) in dimethylformamide as in Example 8b for 2 days (18.5 g, 73%).

C8H6N2O2 (162.1)

NMR (DMSO): δ- 7.1 (d, J-7.5Hz, 1H), 7.41 (s, 1H), 7.59 (d, J-7.5Hz, 1H), 9.44 ( s, 1H), 11.02 (s, 1H), 11.20 (s, 1H) ppm.

d) Preparation of 5- (2,3-Dihiro-2-oxo-1H-benzimidazol-5-yl) -2,4-imidazolidinedione (Compound 20d)

Compound 20c (10.7 g, 0.0659 mole) was treated with 27.8 g (0.289 mole) of ammonium carbonate and 53 g (0.107 mole) of sodium cyanide in methanol / water as described in Example le. Yield: 10 g (65%).

C10H8N4O3 (2323)

NMR (DMSO): δ-5.11 (s, 1H), 6.85 (s, 1H), 6.9 (weak, d, 2H), 8.36 (s, 1H), 10.68 (s) , 3H) ppm.

e) Preparation of DL-α- (tert -butyloxy-carbonyl-amino) -o (2,3-dihydro-2-oxo-1H-benzimidazol-5-yl) -acetic acid (20e)

Compound 20d (8.9 g, 0.0379 mole) was treated with hydrogen bromide (100%, 48%) in analogy to Example 5e. Yield: 10 g (78%).

C ₉ H ₉ N ₃ O ₃ xHBr (336.1).

g (29.7 mmol) of the amino acid hydrobromide in a mixture of 50 ml of 2N sodium hydroxide solution, 50 ml of water and 100 ml of dioxane in analogy to example le) was treated with 19.4 g (89.1 mol) of di-tert-butyl reaction with dicarbonate. Yield: 5.4 g (59%). .

Ct4Ht7N Os ₃ (307.3).

NMR (DMSO): δ-1.4 (s, 9H), 5.08 (d, 1H), 6.99 (s and d, 2H), 7.5 (d, 1H), 10.67 (d) , 2H) ppm.

f) Preparation of D1-7- (2,3-Dihydro-2-oxo-1H-benzimidazol-5-yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid (20Γ)

Compound 20e (1.0 g, 3.25 mmol) in analogy to Example 1f) was treated with triethylamine (0.455 mL, 3.25 mmol) in pivalachloride (0.4 mL, 3.25 mmol) and 3.58 mmol) 7-ACCA Yield: 13 g (71%).

C21H22ClN5O7S (523.9).

1.1 g (2.1 mmol) of Boc-protected cephalosporin are stirred in trifluoroacetic acid / methylene chloride in analogy to Example 1g) and

-17HU 200183Β

Amberlite IRA-68 (acetate) resin was converted to betaine (340 g, 35%).

CitfluCINsOsS x 2 H2O (459.9)

NMR (DCOOD): δ-3.52-4.01 (mm, 2H), 5.25-5.33 (dd, 1H). 5.61 (s, 1H), 5.8-5.9 (dd, 1H), δ

7.39 (d, 2H), 7.51 (s. 1H) ppm.

Preparation of D-7- (2,3-Dihydro-2-oxo-1H-benzimidazole-5-ylcyclylamido) -3-chloro-3-cephem-4-carboxylic acid and its L-10 form

Preparative Separation of Compound (20f) by Liquid Chromatography:

Column: Hibar 250-25, RP-18, 7 µm, 254 nm

The starting material was 925 ml water / 75 ml acetonitrile / 1 ml trifluoroacetic acid

Application: 310 mg

L-shape (vertex I):

Yield: 86 mg (trifluoroacetate)

NMR (DCOOD): δ-3.71 (d, J-18Hz, 1H), 3.95 δ (d, J-18Hz, 1H), 5.31 (d, 1H), 5.61 (s, 1H ), 5.82 (d,

1H), 7.38 (s, 2H), 7.5 (s, 1H) ppm.

HPLC analysis:

Hibar 250-4, RP-8, 10 nm, 254 nm

Mobile phase. 925 ml water / 75 ml acetonitrile / 1 ml trifluoroacetic acid.

Flow: 1.5 ml / min, 0.5 mg / ml.

Retention: 4.5 (content 98%).

D-shape (vertex II):

Yield: 144 mg (trifluoroacetate). 30

HPLC: identical to that given for L-form.

Retention: 5.34 (product content 945%)

NMR (DCOOD): δ - 3.57 (d, 1H), 3.91 (d, 1H), 5.3 (bs, 1H), 5.63 (bs, 1H), 5.92 (bs) s, 1H), 35

7.41 (s, 2H), 755 (d, 1H) ppm.

Example 22

Preparation of D-7- (2-Amino-benzbenzoxazol-5-yl-glycyl-amido) -3-chloro-3-cephem-4-carboxylic acid (I-13)

a) Preparation of D-α-tert-Butyloxy-carbonylamino-α- (3-amino-4-hydroxy-phenyl) -acetic acid (22a)

A solution of 29.1 g (93.2 mmol) of Da- (butyloxycarbonyl) carbonyl-ananone- (3-nitro-4- (hydroxyphenyl) acetic acid) in 300 ml of methanol was added to charcoal. Hydrogenation was carried out under pressure for 2 hours in the presence of 2 g of palladium (10% palladium). The hydrogenated reaction mixture was filtered and the solvent 50 removed in vacuo. The residue was chromatographed on a silica gel column using the following solvent system: methylene chloride / methanol (10: 1), methylene chloride / methanol (1: 1) and methanol. Yield: 9.8 g (37%). 55

C13H18N2O5 (282.3)

NMR (DMSO): δ-1.36 (s, 9H). 4.69 (d, 1H), 6.38 (d, 1H), 6.53-6.58 (s and d, 2H) ppm.

b) Preparation of D-α- (tert-Butyloxy) carbonylamino-α- (2-aminobenzoxazol-5-yl) acetic acid hydrobromide 60 / (22b)

5.5 g (19.5 mmol) of 22a is dissolved in 40 ml of methanol and a solution of 2.2 g (20.5 mmol) of bromocyanane in 20 ml of methanol is added dropwise with stirring at room temperature. the resulting mixture was stirred overnight. The reaction mixture was evaporated to dryness in vacuo. The residue was dissolved in tetrahydrofuran, filtered, mixed with petroleum ether, the precipitated product was filtered off with suction and dried in vacuo. Yield: 4.5 g (57%).

C 14 H 17 N 3 O 5 x HBr XH 2 O (406.2).

NMR (DMSO): δ - 1.36 (s, 9H), 5.18 (d, 1H), 7.19 (d, 1H), 7.34 (s, 1H), 7.47 (d, 1H) ). 7.66 (d. 1H). 8.98 (broad s, 1H) ppm.

c) Preparation of D-7- (2-Aminobenzoxazol-5-yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid

4.5 g (11 mmol) of 22b were treated with 3.1 ml (22 mmol) in analogy to Example 1f). triethylamine, 1.35 mL (11 mmol) of pivaloyl chloride and a solution of 2.84 g (12.1 mmol) in 30 mL of tetrahydrofuran and 12 mL of water in 10% triethylamine solution in THF. Yield: 2.2 g (38%).

1.3 g (2.5 mmol) of Boc group in protected cephalosporin was deprotected and converted to betaine with Amberlite 1RA-68 as described in Example 1g. Yield: 294 g.

C10HmCINsOsS x 2 H2O (459.9)

The crude product was subjected to preparative HPLC (Hibar 250-25, RP).

18.7 μΐη, purification agent: 1000 ml water / 5 ml acetic acid) Yield: 83 mg (product content 99%).

NMR (DCOOD): δ- 3.58 (d, J-18Hz, 1H), 3.91 (d, J-18Hz, 1H), 5.31 (d, J-5Hz, 1H), 7.76 ( d, J = 7.5Hz, 1H), 7.83-7.88 (s and d, 2H) ppm.

Example 23

Preparation of DL-7- (Benzothiazol-5 (6) -yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid (I-14)

a) Preparation of 5 (6) - (Hydroxymethyl) benzthiazole (23a)

55.3 g (0.312 mole) of the methyl ester of benzthiazole carboxylic acid 5 (6) were reacted with DIBAL (20% solution in toluene) in 800 ml (0.936 mole) in 1300 ml of tetrahydrofuran in analogy to Example 1g. g (80%).

C7H7N3O (149.2)

NMR (DMSO): δ 4.71 (s, 2H), 7.44 (d. 1H), 7.83 (s, 1H), 7.92 (d, 1H) ppm.

b) Preparation of bentriazole-5 (6) -carboxaldehyde (23b)

Compound 23a (2.1 g, 14.1 mmol) was mixed with manganese (IV) oxide (0.4 g, 96.6 mmol) in tetrahydrofuran (100 mL) in analogy to Example 1b) at room temperature for 4 days. Yield:

1.4 g (68%).

C7H5N3O (147.1)

NMR (DMSO): δ- 8.0-8.1 (dd, 2H), 8.7 (s, 1H), 10.12 (s, 1H) ppm.

c) Preparation of 5- (Benzothiazol-5 (6) -yl) -2,4-imidazolidinedione (Compound 23c)

Compound 23b (30.9 g, 0.21 mole) was treated with 15.9 g (0.325 mole) of sodium cyanide and 84.7 g (0.882 mole) of anunonium carbonate in a mixture of ethanol and water as described in Example le). Yield: 30 g (66%).

Elemental analysis based on C9H7N5O2 (217.2):

H, 3.25; N, 32.24; topot: C, 49.00; H, 3.60; N, 31.20.

d) Preparation of DL-α-Amino-α (benzthiazol-5 (6) -yl) acetic acid (23d) with 231.7 g (0.508 mol) of barium hydroxide (238 g) (0.138 mol). In 980 ml of water, reflux for 30 hours in analogy to Example 8d. Yield: 16.8 g (63%).

C8H8N4O2 (192.2)

NMR (DCOOD): δ-5.76 (s, 1H), 7.86 (dd. 1H), 8.21 (d. 1H). 8.4 (s, 1H) ppm.

e) Preparation of DL-α- (tert-butyl-oxy) carbonylamino-α (benzthiazol-5 (6) -yl) -acetic acid (23e) g (62.5 mmol) (23d) Compound III was stirred in a mixture of 34 g (156 mmol) of ditert-butyl dicarbonate in dioxane and 2N sodium hydroxide solution overnight as in Example le. The crude product was recrystallized from ethyl acetate / ether / petroleum ether. Yield: 11.9 g (65%).

C13H16N4O4 (292.3)

NMR (DMSO): 6-1.40 (s, 9H), 5.37 (d, 1H), 7.53 (d, 1H), 7.76 (d, 1H), 7.94 (broad, 2H ) ppm.

f) Preparation of DL-7- (Benzothiazol-5 (6) -yl-glycylamido) -3-chloro-3-cephem-4-carboxylic acid

Compound (23e) (4.0 g, 13.7 mmol) in tetrahydrofuran was treated with 1.85 g (13.7 mmol) of 1-hydroxybenzotriazole, 2.82 g (13.7 mmol) of DCC in analogy to Example 10. and 3.22 g (13.7 mmol), 8.5 mL (34.4 mmol) of N, O-bis (trimethylsilyl) acetamide in 7-ACCA in tetrahydrofuran. The dicyclohexylurea is filtered off with suction, the filtrate is evaporated to dryness and the residue is dissolved in water at pH 7. After extraction with ethyl acetate, the aqueous phase was adjusted to pH 2.5 with 2N hydrochloric acid and extracted with two ethyl acetate. The organic layer was washed with neutral, dried over sodium sulfate and evaporated. The residue was precipitated from ethyl acetate / petroleum ether. Yield: 3.8 g (55%).

C20H21CIN6O6S (508.9)

0.8 g (1.57 mmol) of Boc-protected cephalosporin is deprotected and converted to betaine in analogy to Example 1g). Yield: 250 mg (36%).

C15H13ClN6O4S x H2O (444.8)

NMR (DCOOD): δ-3.46-3.82 (mm, 2H). 5.25-5.32 (dd, 1H), 5.76-5.92 (dd and s, 2H), 7.87 (d. 1H), 8.22 (d, 1H), 8.92 (s, H) ppm.

24Lpéld &

D-7- (2-Amino-benzothiazol-5-yl-glycinamide) -3-. chloro-3-cephem-4-carboxylic acid (0-15)

a) Preparation of DL-α-Amino-α- (2-aminobenzothiazol-5-yl) acetic acid (Compound 24a)

5- (2-Aminobenzothlazol-5-yl) 2,4-imidazolidinedione (23.6 g, 0.095 mol) was dissolved in 1000 ml of water, lithium hydroxide (22.7 g, 0.95 mol). solution. Yield: 11.3 g (46%).

C0H9N3O2S x 2 H ₂ O (259.3)

NMR (DMSO): δ-5.71 (s, 1H), 7.96-8.1 (dd. 2H). 8.2 (s. 1H) ppm.

b) Preparation of DL-α- (tert-Butyloxy) carbonylamino-α- (2-aminobenzothiazol-5-yl) acetic acid (24b)

Compound 24a (11.3 g, 0.0436 mole) was stirred with di-tert-butyl dicarbonate (14.3 g, 0.0654 mole, 1.5 equivalents) overnight at room temperature. The crude product was recrystallized from tetrahydrofuran / petroleum ether. Yield: 6.5 g (45%).

For C14H17N3O4S x 1/2 H2O (332.4): C: 50.58, H: 5.5, N: 12.6, S: 9.64%, topot: C: 50.50, H: 5, 2, N: 11ΛS: 9.2%.

NMR (DMSO): δ-1.37 (s, 9H), 5.11 (d, 1H), 7.04 (d, 1H), 7.36 (d, 1H), 7.59 (s, 2H) ), 7.63 (s, 1H) ppm.

c) Preparation of D-7- (2-Aminobenzothiazol-5-yl-glycylamido) -3-doro-3-cephem-4-carboxylic acid

Compound 24b (1.0 g, 3.09 mmol) was treated in analogy to Example 1f with triethylamine (0.433 mL, 3.09 mmol) and pivaloyl chloride (0.38 mL, 3.09 mmol) in dimethyl (10 mL). formate and reacted with 0.726 g (3.09 mmol) of 7-ACCA in triethylamine in a mixture of 7 ml of tetrahydrofuran, 3.5 ml of water and 2 ml of dimethylformamide.

The Boc-protected cephalosporin is deacylated in an analogous manner to that described in Example Ig). Yield: 970 mg (57%).

C16H14CIN5O4S2 x CF3COOH (553.9)

Trifluoroacetate was taken up in 1000 mL of water / 40 mL of acetonitrile / 1 mL of trifluoroacetic acid, a small amount of HP-20 HP (30-60 Mesh, Diaion, Mitsubishi) adsorbent resin was evaporated in vacuo to a crystalline product. The residual material was applied to a column packed with HP-20 resin and eluted with the specified eluent mixture. Yield: peak I (L-form): 115 mg, Ι / Π peak: 383 mg.

Chromatography of the mixture fraction (Peak I / II) on a Whatman column (Magnum 20, 500 x 22.10 µm, CSS-RP-8) with 0.1% trifluoroacetic acid / 1% acetonitrile in water afforded pure D (peak II). which is then converted to betaine with Amberlite IRA-68 (acetate-form) resin. Yield: 48 mg (94% of product by HPLC).

C16H14CIN5O4S2 x 3 H2O (493.9)

NMR (DCOOD): δ- 3.57 (d, J18Hz, 1H), 3.9 (d, J-18Hz, 1H), 5.3 (d, J-5Hz, 1H), 5.73 (s , 1H), 5.91 (d. J-5Hz, 1H), 7.7 (d, 1H), 7.9 (s, 1H), 8.04 (d, 1H) ppm.

Claims (6)

PATENT CLAIMS A process for the preparation of β-tactam compounds of the formula I in the formula X is a group of formula (1), (2) or (3) wherein Hal means halogen, R * is aminobenzothiazolyl, benzimidazolyl, 2-amino-6-methylbenzothiazolyl, aminobenzimidazolyl; trifluoromethylbenzimidazolyl, methylbenzimidazolyl, 2,3-dihydro-2-oxo-1H-benzimidazolyl, aminobenzoxazolyl or benzthiazolyl, characterized in that a compound of the formula (Π) wherein R * -19H 200183Β has the meanings given above and R is an amino-protecting group, after activation by conversion of the carboxyl group to the mixed anhydride, mesylate or activated ester with a compound of formula (X) wherein X is as defined in the disclosure, and then removing the protecting group present. 2. A process according to claim 1, wherein the amino acid of formula (Π) is activated and coupled with the β-octam compound of formula (III) dissolved in an amine salt. 3. A process according to claim 1, wherein the activation is carried out with a pivalic acid chloride or a sulfonic acid derivative of the formula (TV) according to Scheme a or b, wherein R * is as defined in claim 1, R ² is an amino protecting group, T is R ¹³ -SO 2 -O or halogen and R * ³ is 1-optionally substituted with fluoro or chloro, cyano, phenyl, C 1-4 alkyl, C 1-4 alkyl, C 1-4 alkoxy or C 1-4 alkylthio C10 alkyl, or optionally38 with fluorine, chlorine or bromine, cyano, C1-4alkyl, C1-4alkoxy, C1-4alkylthio, C1-4alkyl; phenyl substituted with carbonyl, nitro or trifluoromethyl - via the anhydrides of the formulas Va and Vb. 4. Process according to any one of claims 1 to 3, characterized in that the reaction is carried out at a temperature between -80 ° C and room temperature, preferably between -60 ° C and 0 ° C. 5. A process for the preparation of a pharmaceutical composition having an anti-bacterial effect, characterized in that a compound of the general formula (I) according to claim 1, wherein X, Y, Z, R ¹ , R, R ⁴ R ⁵ , R 1, R ⁸ , R ⁹ and R ¹⁰ are as defined in claim 1, which are converted into pharmaceutical compositions by the customary pharmaceutical excipients. 6. A process for the preparation of animal feeds or animal feed premixes, characterized in that a compound of the formula I is prepared according to claim 1, wherein X, Y, Z, R *. R, R, ^R5, R, ^R7, R, R ⁹ and R ° are as defined in Claim 1 - are mixed with the usual ingredients.